KR20230149808A - Chemical processing systems, instruments and sample cartridges - Google Patents

Abstract

The described embodiments relate to sample cartridges for chemical processing instruments. The sample cartridge includes a primary reaction vessel configured to receive a fluid sample for processing and configured to receive a lid for closing the open top of the primary reaction vessel, and receiving one or more fluid reagents through the open top of the reagent vessel. It includes a reagent container configured to do so. The reagent vessel is connected to the primary reaction vessel through a primary reagent channel with a primary reagent valve disposed in the primary reagent channel to control fluid flow through the primary reagent channel. The sample cartridge is configured to be in fluid communication with the primary reaction vessel when the lid is closed to aspirate the fluid contents of the reagent vessel into the primary reaction vessel and to be connected to a pneumatic module to selectively adjust the pressure within the primary reaction vessel. Additionally includes a pneumatic port. Additional embodiments relate to chemical processing instruments configured to receive a sample cartridge, and systems, methods, and computer-readable media for performing operations using the instrument and the sample cartridge.

Description

Chemical processing systems, instruments and sample cartridges

This application claims priority from the following priority applications, the entire contents of which are incorporated herein by reference: U.S. Provisional Patent Application No. 63/130,450; U.S. Provisional Patent Application No. 63/241,167; U.S. Provisional Patent Application No. 63/292,314; and U.S. Design Patent No. 29/820,394.

technology field

Embodiments generally relate to systems, devices, methods, and computer-readable media for performing operations on a sample, such as nucleic acid extraction operations, as well as sample cartridges for use with a chemical processing instrument.

State-of-the-art automated machines and devices for performing nucleic acid extraction operations, etc. tend to be able to accept and process small sample inputs of less than 1ml, typically about 200ul. However, there are times when it is desirable to extract nucleic acids from a much larger input sample to output sufficient genetic material to perform downstream processing.

Additionally, in standard high-throughput laboratories using well plates and large automated liquid handling robots, bulk samples typically need to be aliquoted into smaller quantities for automated processing. There is, and this is inefficient in using the machine. A typical machine may have many channels, each capable of taking a small sample, but aliquoting reduces the number of patients taking up the machine's run-time.

Another common problem with known automated machines is contamination. The risk is that pipettors are used where containers containing patient-derived material are open or moving within the machine and/or materials that have come into contact with patient-derived material are stored within the device (e.g., pipette tips). It increases every time it happens.

Another known problem with automated machines is the difficulty in controlling the end-to-end process sufficiently to ensure the highest quality.

Another problem is achieving appropriately enriched and/or quantified nucleic acids from the extraction workflow to be directly input into downstream assays. Applications such as MRD typically require high concentrations of DNA (~500ng/μL) that are rarely achieved using available instrumentation for nucleic acid extraction. Additionally, most existing systems aimed at automating cell-free DNA (cfDNA) extractions do not select nucleic acids of a specific size. cfDNA is typically ~150 bp and downstream assays are negatively affected by the presence of genomic DNA (gDNA) contamination in cfDNA samples.

It is desirable to address or improve upon, or at least provide a useful alternative to, one or more deficiencies or shortcomings associated with the prior art for systems, devices, methods and computer-readable media for performing operations on samples. do.

Any discussion of documents, acts, materials, devices, articles, etc. contained herein is not disclosed herein because any or all of such matters form part of the prior art base or existed before the priority date of each appended claim. It should not be regarded as an admission of general general knowledge in the relevant field.

Some embodiments relate to a sample cartridge for a chemical processing instrument, comprising: a primary reaction vessel configured to receive a fluid sample for processing and configured to receive a lid for closing the open top of the primary reaction vessel; a reagent vessel configured to receive one or more fluid reagents through an open top of the reagent vessel, wherein the reagent vessel has a primary reagent valve disposed in the primary reagent channel to control fluid flow through the primary reagent channel; Connected to the primary reaction vessel through the primary reagent channel -; and a primary pneumatic port in fluid communication with the primary reaction vessel and configured to be connected to a pneumatic module to selectively adjust the pressure within the primary reaction vessel when the lid is closed to aspirate the fluid contents of the reagent vessel into the primary reaction vessel. Includes.

The sample cartridge may further include a primary pneumatic channel extending between the primary pneumatic port and the primary reaction vessel, wherein the opening of the primary pneumatic channel to the primary reaction vessel is at some level of the sidewall of the primary reaction vessel. It is located at the top. The opening of the primary pneumatic port to the primary reaction vessel may be located closer to the top of the primary reaction vessel than to the bottom of the primary reaction vessel. In some embodiments, the opening of the primary reagent channel to the primary reaction vessel is located somewhat above the side wall of the primary reaction vessel. In some embodiments, the opening of the primary reagent channel to the primary reaction vessel is located closer to the top of the primary reaction vessel than to the bottom of the primary reaction vessel.

In some embodiments, the sample cartridge may further include a liquid trap configured to restrict passage of liquid out of the sample cartridge. The liquid trap can for example be placed at one end or within the primary pneumatic channel. The liquid trap may be placed at or within the base of the sample cartridge. The liquid trap may include a gas permeable membrane. The liquid trap may comprise a hydrophobic polymer material that can act as a gas-permeable or semi-permeable membrane. A liquid trap may be configured to contain a minimum volume of liquid before clogging or overflowing. The minimum liquid capacity volume of the liquid trap may range, for example, from 1 μL to 1000 μL, 10 μL to 100 μL, 40 μL to 80 μL, or 50 μL to 60 μL.

In some embodiments, the sample cartridge further includes an output vessel configured to receive final output fluid from the primary reaction vessel through a final output channel. The sample cartridge is configured to communicate with the output vessel through the output vessel pneumatic channel and to be connected to a pneumatic module to selectively adjust the pressure of the output vessel to draw final output fluid from the primary reaction vessel to the output vessel through the final output channel. The output container may further include a pneumatic port. The sample cartridge may further include a temporary lid configured to close the output vessel during processing, the temporary lid defining and coupled to an opening for a final output channel and an output vessel pneumatic channel to the output vessel.

In some embodiments, the sample cartridge may be provided without an output vessel and may further include a final output channel configured to convey final output fluid from the primary reaction vessel to the removable output vessel. The cartridge is configured to be in fluid communication with the output vessel through the output vessel pneumatic channel and connected to a pneumatic module to selectively adjust the pressure of the output vessel to draw final output fluid from the primary reaction vessel to the output vessel through the final output channel. It may further include an output container pneumatic port configured to be. The sample cartridge may further include a temporary lid configured to close the output vessel during processing, the temporary lid configured to fluidly connect the final output channel and the output vessel pneumatic channel to the output vessel.

In some embodiments, the sample cartridge includes a quality control vessel configured to receive an aliquot of output fluid for quality control analysis; a quality control channel extending between the quality control vessel and the quality control junction with the final output channel; and a quality control vessel configured to be in fluid communication with the quality control vessel and connected to a pneumatic module to selectively adjust the pressure within the quality control vessel to pull an aliquot of final output fluid from the final output channel through the quality control channel to the quality control vessel. It further includes a management pneumatic port. The quality control vessel can be pre-loaded with dye to be mixed with an aliquot of the final output fluid for quality control analysis.

The sample cartridge includes a buffer container configured to receive buffer through an open top of the buffer container for mixing with the final output fluid for quality control analysis. a buffer channel extending between the buffer channel and the buffer junction with a final output channel between the quality control junction and the primary reaction vessel; and a buffer channel valve disposed in the buffer channel to control the flow of buffer solution through the buffer channel.

The sample cartridge has an intermediate outlet from the final output channel between the quality control joint and the output container; a closed chamber with an open middle outlet; An air-permeable liquid barrier membrane covering the outlet; and an intermediate outlet pneumatic port configured to be in fluid communication with the sealed chamber and connected to the pneumatic module to selectively adjust the pressure within the sealed chamber to draw air from the final output channel through the air-permeable membrane.

In some embodiments, the sample cartridge includes a sealed waste container configured to receive waste liquid from the primary reaction vessel through a waste channel; and a waste pneumatic port in fluid communication with the waste vessel and configured to be connected to a pneumatic module to selectively adjust pressure within the waste vessel to draw fluid from the primary reaction vessel through the waste channel to the waste vessel.

In some embodiments, the sample cartridge is configured to receive primary output fluid from the primary reaction vessel through a primary output channel fluidly connecting the primary reaction vessel to the secondary reaction vessel and connecting the reagent vessel to the secondary reaction vessel. a secondary reaction vessel configured to receive one or more fluid reagents from the reagent vessel through a fluidly connecting secondary reagent channel; a primary outlet valve disposed in the primary outlet channel to control flow through the primary outlet channel; and a secondary reagent valve disposed in the secondary reagent channel to control flow through the secondary reagent channel. The secondary reaction vessel may be sealed, and in some embodiments, the sample cartridge is in fluid communication with the secondary reaction vessel and is connected to the secondary reaction vessel to draw fluid from the primary outlet channel or secondary reagent channel to the secondary reaction vessel. It further includes a secondary pneumatic port configured to be connected to the pneumatic module to selectively adjust the pressure. The sample cartridge may further include a secondary pneumatic channel extending between the secondary pneumatic port and the secondary reaction vessel, wherein an opening of the secondary pneumatic channel into the secondary reaction vessel is lower than the bottom of the secondary reaction vessel. It is located on the upper part of the side wall of the secondary reaction vessel closer to the top of the vessel. The inlet or inlets of the primary output channel and the secondary reagent channel may open into the secondary reaction vessel above the sidewall of the secondary reaction vessel closer to the top of the secondary reaction vessel than the bottom of the secondary reaction vessel.

In some embodiments, in the quality control joint, the quality control channel forms an obtuse angle with a portion of the final output channel extending between the quality control joint and the buffer joint.

In some embodiments, in the buffer joint, the buffer channel forms an obtuse angle with a portion of the final output channel extending between the quality control joint and the buffer joint.

In some embodiments, the pre-cushioned joint portion of the final output channel at the buffer joint forms an obtuse angle with the portion of the final output channel extending between the quality control joint and the buffer joint.

In some embodiments, in the quality control joint, a post-QC joint portion of the final output channel forms an obtuse angle with a portion of the final output channel extending between the quality control joint and the buffer joint.

Some embodiments relate to a chemical processing device configured to receive a sample cartridge according to any of the described embodiments, the device comprising: a reagent dispenser configured to dispense one or more fluid reagents to a reagent container through an open top of the reagent container; and pneumatics connected to the primary pneumatic port of the primary reaction vessel and configured to selectively adjust the pressure within the primary reaction vessel when the lid is closed to draw fluid from the reagent vessel through the primary reagent channel into the primary reaction vessel. Includes modules.

The pneumatic module may be further configured to connect to the output vessel pneumatic port and selectively adjust the pressure of the output vessel to draw final output fluid from the primary reaction vessel to the output vessel through the final output channel. The pneumatic module may be further configured to connect to the quality control pneumatic port and selectively adjust the pressure within the quality control vessel to draw an aliquot of final output fluid from the final output channel through the quality control channel to the quality control vessel. The reagent module may be configured to dispense buffer into a buffer container. The pneumatic module may be further configured to connect to the intermediate outlet pneumatic port and selectively adjust the pressure within the sealed chamber to draw air through the air-permeable membrane from the final output channel. The pneumatic module may be further configured to connect to the waste pneumatic port and selectively adjust the pressure within the waste vessel to draw fluid from the primary reaction vessel to the waste vessel through the waste channel. The pneumatic module may be further configured to connect to the secondary pneumatic port and selectively adjust the pressure of the secondary reaction vessel to draw fluid from the primary outlet channel or secondary reagent channel to the secondary reaction vessel.

The pneumatic module may be configured to sense changes in pressure and/or flow rate to determine when the liquid delivery task is complete. For example, when pressure is adjusted to pull a liquid from one chamber to another, if all the liquid is drawn through the delivery channel, air is drawn along the liquid, resulting in less pressure difference and thus higher flow rate. These changes in pressure and/or flow can be sensed by the pneumatic module and used as a signal to stop pressure operation once the transfer process is complete.

The pneumatic module may be configured to move liquid between the various containers in the cartridge using positive or negative pressure. That is, positive pressure (above atmospheric pressure) is applied to one container to force liquid through a delivery channel into another container, or negative pressure (below atmospheric pressure) is applied to one container to draw liquid through a delivery channel into another container.

In some embodiments, a pneumatic module may be configured to operate using a single pressure level selectively applied to various pneumatic ports at different times to effect different operations. In some embodiments, a pneumatic module may be configured to operate using only two pressure levels that are selectively applied to various pneumatic ports at different times to effect different operations.

In some embodiments, the device may further include an optical module configured to measure a characteristic of a quantity of output fluid contained in a quality control vessel.

The device may be configured to receive a plurality of sample cartridges. The pneumatic module may be configured to be connected to all pneumatic ports of a plurality of sample cartridges and selectively apply pressure to a selected one of the pneumatic ports at a selected time. The reagent module is configured to dispense reagents to each of the plurality of sample cartridges at selected times.

In some embodiments, the device further includes a mechanism and an actuator configured to move the reagent module to various positions within the device, each position having a plurality of actuators to enable the reagent module to dispense one or more reagents to each sample cartridge. Corresponds to each of the sample cartridges.

Some embodiments relate to a chemical processing device configured to receive one or more sample cartridges each containing a fluid sample for processing, each sample cartridge configured to receive a fluid sample for processing and an open top of a primary reaction vessel. a primary reaction vessel configured to receive a lid for closing; a reagent vessel configured to receive one or more fluid reagents through an open top of the reagent vessel, wherein the reagent vessel has a primary reagent channel with a reagent valve disposed in the reagent channel to control fluid flow through the primary reagent channel. Connected to the primary reaction vessel via -; and defining a pneumatic port in fluid communication with the primary reaction vessel; The chemical processing device includes a reagent dispenser configured to dispense one or more fluid reagents into a reagent vessel through an open top of the reagent vessel; and a pneumatic module connected to a pneumatic port of the primary reaction vessel and configured to selectively adjust the pressure within the primary reaction vessel when the lid is closed to aspirate the fluid contents of the reagent vessel into the primary reaction vessel.

Some embodiments relate to a chemical treatment system, comprising: an apparatus according to any of the described embodiments; and one or more of sample cartridges according to any one of the described embodiments.

Some embodiments relate to a chemical processing system comprising one or more sample cartridges, wherein each sample cartridge: is configured to receive a fluid sample for processing and includes a lid for closing the open top of a primary reaction vessel. A primary reaction vessel configured to: a reagent vessel configured to receive one or more fluid reagents through an open top of the reagent vessel, wherein the reagent vessel has a primary reagent channel with a reagent valve disposed in the reagent channel to control fluid flow through the primary reagent channel. Connected to the primary reaction vessel via -; and defining a pneumatic port in fluid communication with the primary reaction vessel; and a reagent dispenser configured to dispense one or more fluid reagents to the reagent vessel through the open top of the reagent vessel. and a pneumatic module connected to a pneumatic port of the primary reaction vessel and configured to selectively adjust the pressure within the primary reaction vessel when the lid is closed to aspirate the fluid contents of the reagent vessel into the primary reaction vessel.

Some embodiments include a method of operating a chemical processing instrument system according to any of the described embodiments, comprising one or more sample cartridges, each containing a fluid sample in a primary reaction vessel, according to any of the described embodiments. In relation to this, the method includes: connecting a pneumatic module to the primary pneumatic port of each sample cartridge; operating the reagent module to dispense one or more reagents to the reagent container of each sample cartridge; Reducing the pressure of the primary reaction vessel of each sample cartridge by operating a pneumatic module to draw the fluid contents of the reagent vessel through each primary reagent channel into the primary reaction vessel of each sample cartridge.

The method may further include operating a shaker on the device to facilitate mixing of the fluids in the primary reaction vessel of each sample cartridge. The method may further include operating the heater of the device to heat the primary reaction vessel of each sample cartridge to a predetermined temperature for a predetermined time.

In some embodiments, the reagent in the primary reaction vessel of each sample cartridge includes functionalized magnetic beads, and the method includes actuating or moving a magnet to maintain the magnetic beads at a selected location within the primary reaction vessel. Includes more.

The method may further include connecting a pneumatic module to the waste pneumatic port of each sample cartridge and reducing the pressure within the waste vessel to draw fluid from the primary reaction vessel to the waste vessel through the waste channel. The method may further include connecting a pneumatic module to the secondary pneumatic port of each sample cartridge and reducing the pressure in the secondary reaction vessel to draw fluid from the primary outlet channel into the secondary reaction vessel. The method may further include connecting a pneumatic module to a secondary pneumatic port of each sample cartridge and reducing the pressure in the secondary reaction vessel to draw fluid from the secondary reagent channel into the secondary reaction vessel.

The method may further include operating a shaker on the device to facilitate mixing of fluids in the secondary reaction vessel of each sample cartridge. The method may further include operating the heater of the device to heat the secondary reaction vessel of each sample cartridge to a predetermined temperature for a predetermined time.

In some embodiments, the reagents in the secondary reaction vessel of each sample cartridge include functionalized magnetic beads, and the method further comprises moving or moving a magnet to secure the magnetic beads at a selected location within the secondary reaction vessel. do.

The method includes connecting a pneumatic module to the waste pneumatic port of each sample cartridge and a waste channel to draw fluid from the secondary reaction vessel to the waste vessel through a secondary waste channel extending between the secondary reaction vessel and the waste vessel. The step of reducing the pressure within the vessel may be further included. The method may further include connecting a pneumatic module to the output vessel pneumatic port of the or each sample cartridge and reducing the pressure in the output vessel to pull the treated fluid from the primary reaction vessel into the output vessel through the final output channel. You can.

In some embodiments, the treated fluid withdrawn from the primary reaction vessel of each sample cartridge is drawn into a secondary reaction vessel and treated with additional reagents before being withdrawn through a final output channel to a final output vessel.

The method comprises connecting a pneumatic module to the quality control pneumatic port of each sample cartridge, and reducing the pressure in the output vessel to draw the treated fluid into the output vessel, through the quality control channel, and at the final output channel. The step may further include reducing the pressure within the quality control vessel to draw an aliquot of treated fluid into the vessel.

The method includes operating a reagent module to inject a buffer solution into the buffer container of the or each sample cartridge; and to draw buffer from the buffer vessel through the buffer channel into the quality control vessel along with a portion of the processed fluid through the final output channel and the quality control channel, prior to lowering the pressure in the quality control vessel of each sample cartridge. It further includes the step of opening the buffer channel valve.

The method includes connecting a pneumatic module to the middle outlet pneumatic port of each sample cartridge and lowering the pressure within the outlet chamber of each sample cartridge before lowering the pressure in the quality control vessel to draw air through an air-permeable membrane in the final output channel. Additional steps may be included.

The method may further include operating the optical module to measure properties of a quantity of treated fluid contained in the quality control vessel of each sample cartridge.

In some embodiments, operating the reagent module to dispense a reagent to the reagent vessel further includes operating mechanisms and actuators to move the reagent module to various locations within the device, each location containing one or more sample cartridges. Correspond to each.

Some embodiments relate to a computer-readable storage medium storing instructions that, when executed by a computer, cause the computer to perform any one of the described methods.

Some embodiments relate to methods of using the system of any of the described embodiments, the method comprising: placing a fluid sample into a primary reaction vessel of each sample cartridge; applying a lid to seal and close the open top of the primary reaction vessel of said or each sample cartridge; Inserting each sample cartridge into the corresponding cartridge slot of the instrument; and operating the device to process the fluid sample.

Some embodiments relate to a method of using the system of any one of the described embodiments, the method comprising: placing a fluid sample into a primary reaction vessel of each sample cartridge; applying a lid to seal and close the open top of the primary reaction vessel of said or each sample cartridge; Inserting each sample cartridge into the corresponding cartridge slot of the instrument; and operating the device to process the fluid sample.

The method may further include removing each sample cartridge from the device once the fluid sample has been processed. The method may further include removing the output container containing the processed fluid sample from the sample cartridge. The method may further include removing the temporary lid from the output container.

Some embodiments relate to a sample cartridge for use with a fluid analysis device, the cartridge comprising: a sample container configured to receive a fluid sample for analysis; a buffer container configured to contain a buffer solution; an assay vessel configured to contain a mixed fluid comprising an aliquot of a fluid sample mixed with at least a portion of an assay buffer; a sample channel extending between the sample vessel and the first junction; a sample channel valve disposed in the sample channel to control the flow of sample through the sample channel; a buffer channel extending between the buffer container and the first junction; a buffer channel valve disposed in the buffer channel to control the flow of buffer solution through the buffer channel; a metering channel in fluid communication with the buffer channel and the sample channel, wherein the metering channel extends between the first junction and the second junction; an assay vessel channel in fluid communication with the metering channel and extending between the second junction and the assay vessel; and an assay vessel pneumatic port in communication with the assay vessel and configured to be connected to a pneumatic module to selectively adjust the pressure of the assay vessel to draw fluid into the assay vessel through the assay vessel channel.

In some embodiments, at least one of the sample channel valve and the buffer channel valve allows a portion of the fluid sample to be aspirated into the metering channel and allows buffer solution to flow through the buffer channel, the metering channel, and the assay vessel channel with an aliquot of the fluid sample for analysis. It includes an active valve that can be selectively opened and closed to allow inflow into the analysis vessel. The assay vessel may be pre-loaded with a dye configured to mix with the buffer and fluid sample to facilitate analysis.

The sample cartridge includes an intermediate outlet in fluid communication with the metering channel through a second junction; an outlet chamber in which the middle outlet is open; A liquid-permeable liquid barrier membrane covering the outlet; and an intermediate outlet pneumatic port in fluid communication with the outlet chamber and configured to be connected to a pneumatic module to selectively adjust the pressure of the outlet chamber to draw air from the metering channel through the breathable membrane, wherein the intermediate outlet is configured to be connected to the sample. The arrangement is such that liquid entering the metering channel from the channel or buffer channel is allowed to fill the metering channel but is not allowed to proceed to the analysis vessel channel. The intermediate outlet may be located at the second junction.

In some embodiments, the sample cartridge further includes an outlet channel extending between the second junction and the outlet, such that liquid entering the metering channel from the sample channel or buffer channel can fill the metering channel and enter the outlet channel but not the analytical vessel channel. Do not allow entry.

The sample channel includes an output vessel in fluid communication with the metering channel through the second junction and through the output channel; and an output vessel pneumatic port configured to communicate with the output vessel and be connected to the pneumatic module to selectively adjust the pressure of the output vessel to draw fluid from the metering channel to the discharge vessel through the second junction and the output channel. . The output channel may extend from the second junction to the output container. The output channel may extend between the intermediate outlet and the output container.

In some embodiments, the buffer channel valve includes a pressure actuated valve including a buffer channel valve pneumatic port configured to be coupled to a pneumatic module to selectively open or close the buffer channel valve.

Some embodiments relate to a fluid analysis instrument configured to receive a sample cartridge of any one of the described embodiments, wherein the device: connects to an assay vessel pneumatic port and draws fluid into the assay vessel through the assay vessel channel; a pneumatic module configured to selectively adjust the pressure of the analytical vessel; and an analysis module configured to measure properties of the fluid in the analysis vessel.

Some embodiments relate to a fluid analysis device comprising a sample cartridge according to any one of the described embodiments, the device being: connected to an analysis vessel pneumatic port and configured to draw fluid into the analysis vessel through an analysis vessel channel; a pneumatic module configured to selectively adjust the pressure of the analytical vessel; and an analysis module configured to measure properties of the fluid in the analysis vessel.

The analysis module may include an optical source configured to illuminate the fluid within the analysis vessel and an optical detector configured to detect or measure light transmitted from the fluid within the analysis vessel.

In some embodiments, the pneumatic module is further connected or configured to be connected to the intermediate outlet pneumatic port and configured to selectively adjust the pressure in the outlet chamber to draw air from the metering channel through the air-permeable membrane. The pneumatic module may be further connected or configured to be connected to the output vessel pneumatic port and may be configured to selectively adjust the pressure of the output vessel to draw fluid from the metering channel through the second junction and the output channel to the output vessel. . The pneumatic module may be additionally connected or configured to be connected to the buffer channel valve pneumatic port and configured to selectively open and close the buffer channel valve.

In some embodiments, the device is configured to receive a plurality of sample cartridges containing a fluid sample of any of the described embodiments.

The device may further include a mechanism and an actuator configured to move the analysis module to various positions corresponding to each of the sample cartridges for analysis of the fluid in the analysis vessel of each sample cartridge.

Some embodiments relate to a fluid analysis system, comprising: any one of the described embodiments; and one or more of the sample cartridges of any one of the described embodiments.

Some embodiments relate to a method of operating a fluid analysis device of any of the described embodiments containing a fluid sample in a sample vessel, the method comprising: flowing the sample fluid through the sample channel without the sample fluid proceeding into the analysis vessel channel. operating the pneumatic module to draw the metering channel from the sample vessel to the second junction; and operating the pneumatic module to draw fluid from the buffer container through the buffer channel, metering channel, and analysis channel into the analysis vessel along with an aliquot of sample fluid in the metering channel.

The method includes operating a pneumatic module to reduce the pressure in the assay vessel for a predetermined period of time to pull the sample fluid through the sample channel to the second junction from the sample channel to the metering channel without the sample fluid advancing into the assay vessel channel. ; and operating the pneumatic module to reduce the pressure within the assay vessel after a predetermined period of time to draw fluid from the buffer vessel through the buffer channel, measurement channel, and analysis channel along with an aliquot of sample fluid from the metering channel into the analysis vessel. It may further include.

In some embodiments, the method includes reducing the pressure at the intermediate outlet by operating a pneumatic module to draw sample fluid from the sample vessel through the sample channel and into the metering channel until the sample fluid encounters an air permeable barrier; and thereafter operating a pneumatic module to draw fluid from the buffer vessel through the buffer channel, metering channel and analysis channel into the analysis vessel along with an aliquot of sample fluid to reduce the pressure in the analysis vessel. You can.

The method includes operating a pneumatic module to reduce the pressure in the output vessel for a predetermined period of time to draw the sample fluid into the metering channel from the sample vessel to the second junction through the sample channel without the sample fluid proceeding into the analysis vessel channel. ; and operating a pneumatic module to reduce the pressure in the assay vessel after a predetermined period of time to draw fluid from the buffer vessel through the buffer channel, metering channel, and analysis channel into the analysis vessel along with an aliquot of sample fluid from the metering channel. It may further include. In some embodiments, the operation of the pneumatic module to draw fluid from the buffer container through the buffer channel continues until the metering channel is filled with air. The method may further include subsequently operating a pneumatic module to reduce the pressure of the output vessel to draw sample fluid from the sample vessel into the output vessel.

In some embodiments, the method includes operating a pneumatic module to keep the buffer valve closed while drawing fluid from the sample vessel; and then operating the pneumatic module to keep the buffer valve open so that fluid can be withdrawn from the buffer container.

Some embodiments relate to a method of operating a fluid analysis device of any of the described embodiments, comprising receiving one or more of the sample cartridges of any of the described embodiments, wherein the sample container of each sample cartridge includes a fluid sample. and the method includes: operating the pneumatic module to draw the sample fluid from the sample channel or the sample container of each sample cartridge through the sample channel to the metering channel to the second junction without advancing the sample fluid into the analysis vessel channel; and then operating the pneumatic module to draw fluid from the buffer container of each sample cartridge through the buffer channel, metering channel, and analysis channel into the analysis vessel along with an aliquot of sample fluid in the metering channel.

In some embodiments, the method includes connecting a pneumatic module to the assay vessel pneumatic port of each sample cartridge; Pressure is maintained in the assay vessel of the or each sample cartridge for a predetermined period of time by operating a pneumatic module to draw sample fluid into the metering channel from the sample vessel to the second junction through the sample channel without the sample fluid advancing into the assay vessel channel. reducing step; and to reduce the pressure in the assay vessel of said or each sample cartridge after a predetermined period of time to draw fluid from the buffer vessel through the buffer channel, metering channel and analysis channel into the analysis vessel along with an aliquot of sample fluid from the metering channel. It further includes operating the pneumatic module.

The method includes connecting a pneumatic module to an analysis vessel pneumatic port and a mid-outlet pneumatic port of the or each sample cartridge; activating a pneumatic module to reduce the pressure at the intermediate outlet of each sample cartridge to draw sample fluid from the sample vessel through the sample channel and into the metering channel until the sample fluid encounters an air permeable barrier; and thereafter operating a pneumatic module to draw fluid from the buffer vessel through the buffer channel, metering channel, and analysis channel into the analysis vessel along with an aliquot of sample fluid to reduce the pressure in the analysis vessel of each sample cartridge. It may further include.

The method includes connecting a pneumatic module to an analysis vessel pneumatic port and an output vessel pneumatic port of the or each sample cartridge; Pressure is maintained in the output vessel of said or each sample cartridge for a predetermined period of time by operating a pneumatic module to draw sample fluid from the sample vessel to the second junction through the sample channel and into the metering channel without the sample fluid advancing into the analysis vessel channel. reducing step; and pneumatically to reduce pressure in the assay vessel of said or each sample cartridge after a predetermined period of time to draw fluid from the buffer vessel through the buffer channel, metering channel and analysis channel into the analysis vessel along with an aliquot of sample fluid from the metering channel. The step of operating the module may further be included.

The operation of the pneumatic module to draw fluid from the buffer container through the buffer channel may continue until the metering channel of each sample cartridge is filled with air.

In some embodiments, the method includes connecting a pneumatic module to the output vessel pneumatic port of each sample cartridge; and drawing a buffer solution into the analysis vessel to draw sample fluid from the sample vessel into the output vessel, and then operating a pneumatic module to reduce the pressure in the output vessel of each sample cartridge. The method includes connecting a pneumatic module to the buffer valve pneumatic port of each sample cartridge; operating a pneumatic module to maintain the buffer valve of each sample cartridge in a closed state during the period during which fluid is extracted from the sample vessel; and then operating the pneumatic module to hold the buffer valve of each sample cartridge open to allow fluid to drain from the buffer container.

The method may further include operating the analysis module to subsequently measure properties of the fluid within the analysis vessel.

The method may further include transmitting data regarding the measured attribute to an external computing device.

Some embodiments relate to a computer-readable storage medium storing instructions that, when executed by a computer, cause the computer to perform any one of the described methods.

Some embodiments relate to methods of using any of the described systems, comprising: placing a fluid sample into a sample container of the or each sample cartridge; Inserting each sample cartridge into the corresponding cartridge slot of the instrument; and operating the device to analyze the fluid sample. The method may further include removing each sample cartridge from the device once the fluid sample has been processed.

Some embodiments relate to a sample cartridge for use with a fluid analysis device, the cartridge comprising: a sample container configured to receive a fluid sample for analysis; a buffer container configured to contain a buffer solution; a sealed assay vessel configured to contain a mixed fluid comprising an aliquot of a fluid sample mixed with at least a portion of an assay buffer; a first channel extending between the sample vessel and the analysis vessel; a second channel extending from the buffer container to a junction with the first channel; a first valve disposed in the first channel between the sample vessel and the junction; a second valve disposed in the second channel between the buffer container and the joint; and a pneumatic port in communication with the analysis vessel and configured to be connected to a vacuum pump for aspirating fluid from the first channel to the analysis vessel, wherein at least one of the first valve and the second valve directs a quantity of the fluid sample into the first channel. and optionally open and opened to allow passage of the junction and then allow the buffer to flow through the second channel past the junction into the first channel to carry and mix a portion of the fluid sample and then flow into the analytical vessel for analysis. Contains an active valve that can be closed.

Some embodiments relate to a chemical processing device configured to receive a sample cartridge containing a fluid sample volume of at least 0.2 mL, wherein the device is operated according to instructions stored on a computer-readable storage medium to detect contamination of the device or another sample. Processing the samples while keeping them isolated to prevent cross-contamination with; Selecting nucleic acids using specific chemistry, incubation conditions, bead selection, and elution parameters; selecting a desired range of single or double stranded nucleic acid sizes in the processed fluid product and discarding unwanted material outside the desired range; increasing the concentration of the selected nucleic acid product; and processing an aliquot of the treated fluid product, mixing it with a specific fluorochrome for the selected nucleic acid, and quantifying the properties of the product, e.g., quantification relative to a standard reference curve or a calibrated reference curve. configured to perform any two or more of the steps.

Some embodiments relate to a kit comprising a sample cartridge according to any one of the described embodiments, and a temporary lid configured to close the output vessel during processing, wherein the temporary lid outputs a final output channel and an output vessel pneumatic channel. It is configured to be fluidly connected to the container. The kit may further include an output container, for example an Eppendorf tube.

The temporary lid may be configured to be mechanically coupled to the cartridge by a resiliently flexible body. The main body may be formed integrally with the temporary lid. The body may be configured to press the output container against the cartridge when connected. The main body can define a channel for fluidly connecting the final output channel and the output container pneumatic channel to the output container.

Throughout this specification, the word "comprise" or variations such as "comprises" or "comprising" refers to a specified element, integer, or step, or to a group of elements, integers, or steps. It will be understood to mean including a group, but not excluding another element, integer or step or group of elements, integers or steps.

The various drawings of the accompanying drawings merely illustrate exemplary embodiments of the present disclosure and should not be considered limiting its scope.
1A is a schematic diagram of an instrument configured to receive one or more sample cartridges each containing a sample for processing according to some embodiments;
Figure 1B is a perspective view of the device of Figure 1A according to some embodiments;
FIG. 1C is a cutaway perspective view illustrating some internal components of the device of FIG. 1A according to some embodiments;
Figure 2A is a perspective view of a sample cartridge for use with an instrument according to some embodiments;
Figure 2b is a top view of the sample cartridge of Figure 2a;
Figure 2C is a side view of the sample cartridge of Figure 2A;
Figure 2D is a bottom view of the sample cartridge of Figure 2A;
Figure 2E is a bottom view of the sample cartridge of Figure 2A illustrating further details;
Figure 2F is a bottom view of a fluid metering arrangement according to some embodiments that may form part of the sample cartridge of Figure 2A;
Figure 2G is a circuit diagram of the sample cartridge of Figure 2A according to some embodiments;
Figures 2H and 2I are perspective views showing alternative channel arrangements for the sample cartridge of Figure 2A according to some embodiments;
Figures 2J-2N illustrate alternative valves for the sample cartridge of Figure 2A according to some embodiments;
Figure 2O is an enlarged perspective view of the middle outlet of a sample cartridge according to some embodiments;
Figure 3A is a perspective view of a reagent module of the instrument of Figures 1A-1C according to some embodiments;
Figure 3b is a perspective view of the reagent cartridge of the reagent module of Figure 3a;
Figures 4A and 4B are schematic diagrams of the optics module of the device of Figures 1A-1C according to some embodiments;
Figure 5A is a schematic diagram of the device of Figures 1A-1C showing components of a pneumatic module, a thermal module, a magnetic module, a mixing module, and a motion module according to some embodiments;
Figure 5B is a side view of the device of Figures 1A-1V showing components of a thermal module, magnetic module, mixing module, and motion module according to some embodiments; and
Figure 5C is a perspective view of the core unit of a device according to some embodiments;
Figure 5D is a close-up perspective view of the pneumatic interface plate of the core unit of Figure 5C;
Figure 5e shows the internal components of the core unit with other components omitted for clarity;
Figure 5F is a perspective view of the sample cartridge of Figure 7A installed in a socket of the core unit of Figure 5C and a heating assembly engaging the sample cartridge;
Figure 6 is a schematic diagram of a control module of the device of Figures 1A-1C according to some embodiments;
Figure 7A is a perspective view of a sample cartridge according to some embodiments;
Figure 7b is a bottom perspective view of the sample cartridge of Figure 7a;
Figure 7c is an exploded view of the sample cartridge of Figure 7a;
Figure 7D is a close-up exploded view of the base and pneumatic channel plate of the sample cartridge of Figure 7A;
Figure 7E is a bottom perspective view of the base and pneumatic channel plate of the sample cartridge of Figure 7A;
Figure 7F is a bottom cross-sectional view of the top portion of the base of the sample cartridge of Figure 7A;
Figure 7g is a bottom view of the base of the sample cartridge of Figure 7a;
Figure 7h is a bottom view of the base membrane of the sample cartridge of Figure 7a;
Figure 7i is a bottom view of the PSA layer of the sample cartridge of Figure 7a;
Figure 7J is a bottom cross-sectional view of the upper portion of the pneumatic channel plate of the sample cartridge of Figure 7A;
Figure 7K is a bottom view of the pneumatic channel plate of the sample cartridge of Figure 7A;
Figure 7l is a superimposed view of Figures 7h and 7j;
Figure 7m is a vertical cross-sectional view of the sample cartridge portion of Figure 7a;
Figure 7n is a cross-sectional view of the bottom portion of the primary reaction vessel of the sample cartridge of Figure 7a;
Figure 7O is an enlarged perspective cross-sectional view of the upper portion of the primary reaction vessel of the sample cartridge of Figure 7A;
Figure 7P is a cross-sectional view of the primary reaction vessel lid of the sample cartridge of Figure 7A;
Figure 7q is an enlarged view of the secondary reaction vessel shown in Figure 7m;
Figure 7r is a perspective view of another upper portion of the secondary reaction vessel;
Figure 7s is an enlarged assembled view of the sample cartridge portion of Figure 7a;
Figure 7T is a perspective view of the waste fluid trap and primary fluid trap of the sample cartridge of Figure 7A;
Figure 7u is a perspective view of the secondary fluid trap of the sample cartridge of Figure 7a;
Figure 7v is an enlarged assembled view of another portion of the sample cartridge of Figure 7a;
Figure 7W is an enlarged assembled view of the gas permeable membrane and middle outlet of the sample cartridge of Figure 7A;
Figure 7x is a perspective cross-sectional view of the assembled middle outlet and gas permeable membrane of the sample cartridge of Figure 7a;
Figure 7y is a bottom view of the weld pattern to the base membrane of the sample cartridge of Figure 7a;
Figure 7z is an enlarged bottom view of the metering channel and microfluidic junction of the sample cartridge of Figure 7a;
Figure 8A is a perspective view of a fluid transfer device according to some embodiments;
Figure 8b is a perspective view of the fluid delivery device of Figure 8a installed on the sample cartridge of Figure 7a; and
Figure 8C is an enlarged cross-sectional perspective view of the connection between the fluid transfer device and the sample cartridge.

Embodiments generally relate to systems, devices, methods, and computer-readable media for performing operations on samples, such as nucleic acid extraction operations.

There are many chemical process workflows that typically involve several steps requiring the transfer of fluid samples to different vessels and/or different devices. At each transfer step there is the potential for sample spillage, contamination of instruments with the sample, and potential cross-contamination with other samples for processing.

Some embodiments relate to a sample cartridge for a chemical processing instrument that facilitates a workflow process to isolate samples and mitigate cross-contamination. The sample cartridge includes a primary reaction vessel and a reagent vessel arranged to receive a lid. The primary reaction vessel and the reagent vessel are connected or in fluid communication through a primary reagent channel. The primary reagent channel may have a primary reagent valve disposed in the primary reagent channel to control fluid flow through the primary reagent channel. The primary pneumatic port is configured to be in fluid communication with the primary reaction vessel and connected to the pneumatic module. The liquid contents of the reagent vessel can be drawn into the primary reaction vessel by using a pneumatic module to selectively adjust the pressure within the primary reaction vessel when the lid is closed.

The sample cartridge may further include a primary pneumatic channel extending between the primary pneumatic port and the primary reaction vessel, wherein the opening of the primary pneumatic channel to the primary reaction vessel is above a portion of a side wall of the primary reaction vessel. is located in The opening of the primary pneumatic port to the primary reaction vessel may be located closer to the top of the primary reaction vessel than to the bottom of the primary reaction vessel. This reduces the chance of liquid specimens aspirating into the pneumatic module and potentially contaminating the instrument.

In some embodiments, the opening of the primary reagent channel to the primary reaction vessel is located partially above a side wall of the primary reaction vessel. In some embodiments, the opening of the primary reagent channel to the primary reaction vessel is located closer to the top of the primary reaction vessel than to the bottom of the primary reaction vessel. This may reduce the likelihood of liquid specimens entering the reagent channel and subsequently the reagent vessel, which could otherwise lead to cross-contamination in the instrument.

Various other features in accordance with some embodiments are described below, including sealed containers for processing of samples, separate open containers for holding reagents, one-way valves, and fluids with which reaction vessels can be sealed. Cross-contamination is further mitigated by including a pneumatic system to drive the flow.

For example, there are many chemical processing workflows that require quantitation of an aliquot of a fluid sample for analysis or measurement of one or more properties of the fluid sample, such as quality control.

Some embodiments relate to an arrangement of channels and valves that allow accurate fluid metering to isolate aliquots of known volumes for analysis or quality control, as described below with respect to FIGS. 2E-2G. will be. This arrangement can be included in a sample cartridge for use in an instrument or dedicated fluid analysis device.

Referring first to FIGS. 1A-1C, a device 100 according to some embodiments is shown. Device 100 may be configured to receive one or more sample cartridges 200, each containing a sample for processing. Instrument 100 may be configured to perform one or more operations on a sample, such as: chemical processing steps, heating, cooling, incubation, mixing, analysis, or measurement. The device 100 may be configured to perform multiple operations on a sample that would typically be performed in separate instruments or manually in a laboratory.

In some embodiments, device 100 may be configured to perform one or more nucleic acid extraction operations on a sample. For example, nucleic acids (e.g., DNA or RNA) are extracted from a patient sample and a concentrated and optionally quantified output fluid containing the nucleic acids from the sample is provided.

Device 100 may include various modules configured to perform operations on a sample. These may include one or more selected from the following groups: reagent module 300, optical module 400, pneumatic module 500, thermal module 600, magnetic module 700, mixing module 800, motion A control module 101 that controls operations performed by the module 900 and the device 100. Device 100 may also have or be connected to a power supply 102 to power various modules.

Reagent module 300 may be configured to dispense selected reagents to sample cartridge 200 . For example, reagent module 300 may include a plurality of repositories each containing a corresponding plurality of reagents for use in the operations of instrument workflows. Reagent module 300 may include one or more pumps, channels and dispensing nozzles to selectively dispense a controlled amount of reagent to selected sample cartridges 200 at selected times as part of one or more instrument workflows. .

For example, reagent module 300 may include a syringe pump configured to control the distribution of reagents. The reagent module may include two dispensing nozzles configured to dispense different reagents at different times. Previous reagents can be flushed from the nozzle before dispensing subsequent reagents into the cartridge. In some embodiments, the device may include a waste receptacle and the reagent module may be configured to dispense some reagents to the waste receptacle to flush previous reagents from the nozzles prior to dispensing into the cartridges.

In some embodiments, reagent module 300 may include a sensor configured to detect the presence or absence of liquid in a portion of the reagent module. For example, the sensor may be configured to monitor the dispensing outlet or dispensing outlet tube to indicate or confirm when reagents are being dispensed through the outlet tube. In some embodiments, multiple sensors may be configured to monitor fluid levels in multiple different fluid lines and/or reagent reservoirs within the reagent module.

One or each sensor may include an optical sensor, such as a light source and a light detector arranged to detect light from a light source that passes through (or reflects) a translucent or transparent wall of the fluid line or reservoir.

A sensor or sensors in the reagent module may be connected to a user interface or indicator LED to, for example, confirm priming of the reagent lines; Determine when reagents are dispensed; Or it can indicate when reagents are exhausted or need to be replaced in the near future.

Tubing used for the reagent lines within the reagent module may include any suitable materials, such as silicone tubing or PTFE tubing, for example. Appropriate dimensions of the tubing may be selected depending on the liquids being dispensed for the particular application. For example, the inner diameter may be approximately 0.3 mm and the outer diameter may be approximately 1.6 mm.

The optical module 400 may include optical sensors or detectors for optical inspection of materials or fluids contained within the sample cartridge 200. The optical module 400 may further include one or more light sources for illuminating materials or fluids included in the sample cartridge 200 for inspection. Optics module 400 detects specific frequencies and/or intensities of light in the optical or near optical spectrum to determine specific properties of materials or fluids contained within the sample cartridges, such as concentration or density, for example. and/or may be configured to measure.

For example, the optical module may include an epifluorescent system comprising a UV LED light source, which transmits light through a band-pass filter, excites fluorescence in a dye within the cartridge, and (excite), causing emission from the dye that is detected by a photodiode.

Pneumatic module 500 may be configured to apply pressure differences to drive fluid flow along specific flow paths of sample cartridge 200 or device 100.

The pneumatic module may be configured to move liquids between the various containers of the cartridge using positive or negative pressure. That is, positive pressure (above atmospheric pressure) is applied to one container to force liquid through delivery channels into another container, or negative pressure (below atmospheric pressure) is applied to one container to pull liquid from another container through delivery channels.

In some embodiments, a pneumatic module may be configured to operate using a single pressure level selectively applied to various pneumatic ports at different times to effect different operations. In some embodiments, a pneumatic module may be configured to operate using only two pressure levels that are selectively applied to various pneumatic ports at different times to effect different operations.

For example, two pressure levels may be needed if the cartridge contains pressure-actuated valves that require a higher pressure than the actuation pressure to transfer liquids through the channels.

Any suitable pressure differential (e.g., vacuum pressure) can be used to induce flow in the cartridge, but too little pressure can result in particularly long liquid transfer times and too large a pressure gradient can cause splashing. ) or sputtering, which may be desirable in certain applications or high shear rates of the liquid flow that could potentially damage certain molecules, for example nucleic acids. You may not. The appropriate vacuum pressure also depends on the viscosity or range of viscosities of the liquids used in a given application. In some embodiments, the actuating vacuum pressure may range, for example, from 50 mBar to 500 mBar, from 80 mBar to 300 mBar, from 100 mBar to 200 mBar, from 100 mBar to 120 mBar, about 100 mBar, or about 120 mBar.

Thermal module 600 may include one or more heating modules configured to control and/or adjust the temperature of the sample cartridge 200 and materials contained therein during different operations of instrument workflows, such as incubation for culturing. Alternatively, it may include cooling elements.

The magnetic module 700 may include one or more permanent magnets or electromagnets configured to control the movement of magnetic beads in the sample cartridge 200. For example, magnetic beads may be used in primary reaction vessel 210 (FIG. 2A) for components of the sample to bind during certain operations of the instrument workflow. Magnetic module 700 may be configured to hold the magnetic beads in place while liquid drains from the magnetic beads.

Alternatively, non-magnetic functionalized beads can be used for binding and a filter can be used to restrict the beads from leaving the reaction vessels. Another alternative is to use a porous material such as a frit with a functionalized surface for bonding and liquids can be drawn through the frit to achieve the desired reactions.

In other embodiments, chemical catalysts or reactants may be provided as coatings on the surface of solid structures, such as beads or porous solids, for reaction with liquids in reaction vessels.

Mixing module 800 may be configured to facilitate mixing of fluids in primary and/or secondary reaction vessels 210, 220 (FIG. 2A) during certain operations of instrument workflows. For example, mixing module 800 may include an orbital shaker, such as an eccentric weight, configured to be rotated by a motor.

Motion module 900 moves certain modules to different positions corresponding to cartridge slots 120 to perform operations on corresponding cartridges 200 (and/or samples therein) at different times. It may include one or more motors or actuators. For example, the reagent module 300 and the optical module 400 can be moved to different cartridge locations and perform operations on corresponding cartridges 200 at those locations.

Control module 101 may include electronic hardware that communicates with other modules of device 100 and software configured to control operations of device modules according to a selected device workflow.

Each of the modules according to some embodiments is described further below.

Device 100 may be configured to connect to an external computer system, such as a laboratory information system 103. Instrument 100 may be configured to transmit data, such as analytical or measurement data associated with a sample in sample cartridge 200, to an external laboratory information system 103. In some embodiments, device 100 may be configured to receive information from an external laboratory information system, such as data about a sample, reference data for comparison, or commands to control the operations of device 100. You can.

In some embodiments, device 100 may include user interface 105. The user interface 105 may include a display of the device 100 itself or an external display that communicates with the device 100. User interface 105 may be configured to allow a user to select a workflow program to perform on samples in sample cartridge 200 . A workflow program may be selected from a list of other programs containing different workflow operations configured to accomplish different processes.

For example, the list of workflow programs may include: DNA (e.g., genomic DNA, rearranged immunoglobulin or TCR DNA, cDNA, cfDNA) and RNA (e.g., mRNA Extraction, isolation, enrichment, enrichment or quantification of natural or non-naturally occurring nucleic acids, including primary RNA transcripts, transfer RNAs or microRNAs. Non-naturally occurring nucleic acids to be isolated include glycol nucleic acids, threose nucleic acids, locked nucleic acids, and peptide nucleic acids. Other workflow programs include the preparation of nucleic acids for amplification (e.g., PCR library preparation) or other types of manipulation, such as sequencing for applications such as in vitro transcription and/or translation, or insertion into vectors. Or it may include analysis.

User interface 105 may also display information regarding samples and/or an indication of the workflow program currently in progress or a particular step of the workflow program.

Device 100 may include a chassis or housing 110 to accommodate some or all modules. In some embodiments, housing 110 may be configured to be stackable with others in device 100 such that multiple devices 100 can be stacked vertically or arranged side-by-side, for example in a laboratory. .

The device 100 may include a plurality of cartridge slots or sockets 120 each configured to receive a corresponding sample cartridge 200. In this way, multiple samples can be processed simultaneously. Cartridge slots 120 may be defined at least in part by external openings of housing 110 configured to receive sample cartridge 200 .

Some modules may have dedicated components for each cartridge socket 120. Some modules may act on all cartridges 200 in cartridge slots 120 simultaneously. Some modules may be configured to act on different cartridges 200 in cartridge slots 120 at different times.

1C, a cross-sectional view of device 100 is shown illustrating a portion of motion module 900 according to some embodiments. A plurality of sample cartridges 200 are shown disposed in corresponding plurality of cartridge slots 120. Cartridges 200 and cartridge slots 120 are arranged in parallel extending across a portion of device 100.

The motion module 900 may include a track 910 extending across a plurality of cartridge slots 120 and a carriage 920 configured to move along the track 910 . Carriage 920 may be configured to carry one or more modules, such as reagent module 300 and optical module 400, and may be configured to perform operations on sample cartridge 200 by moving them to different cartridge locations. there is. An actuator, such as a motor operated by control module 101, may be configured to move carriage 920 between a rest position and various cartridge positions.

In some embodiments, the motion module 900 includes a plurality of carriages 920 and corresponding tracks 910 each configured to carry a different module, such as, for example, a reagent module 300 and an optical module 400. may include.

Track 910 may include motion stages 912, which may include markings or other indicia to allow module(s) to perform operations on selected sample cartridges 200. A plurality of carriage positions may be specified that correspond to cartridge positions that appropriately align the carriage module(s) with the cartridge slots 120 and corresponding sample cartridges 200. Motion module 900 may include one or more sensors disposed on carriage 920 configured to detect an indication signal for carriage 920 to be stopped at a selected carriage position. Alternatively, known actuator states (e.g., angle of a stepper motor) corresponding to specific carriage positions may be selected to move the carriage to the selected carriage position.

2A-2N, a sample cartridge 200 is shown according to some embodiments. The sample cartridge 200 includes a base 202, a primary reaction vessel 210, a reagent vessel 230, and an output vessel 250. Sample cartridge 200 may include various features for various applications depending on the operations performed on the sample.

In some embodiments, sample cartridge 200 may further include an optional secondary reaction vessel 220, as shown in Figure 2A and described further below.

In some embodiments, sample cartridge 200 may further include an optional waste container 240, as shown in FIG. 2A and described further below.

In some embodiments, sample cartridge 200 may further include an optional quality control module 260, as shown in FIG. 2A and described further below.

Sample cartridge 200 includes various vessels (primary reaction vessel 210, reagent vessel 230 and output vessel 250, an optional secondary reaction vessel 220 in some embodiments, and an optional waste vessel ( 240), defining channels connecting the optional quality control module 260) so that the vessels are in fluid communication and fluids (including liquids and potentially liquid slurries containing solids) are exchanged between the vessels. It can be done as much as possible. Sample cartridge 200 may include valves that can control fluid exchange between the vessels by selectively allowing or disallowing flow through the channels. The network of valves and channels is described further below according to some embodiments.

In some embodiments, vessels (primary reaction vessel 210, reagent vessel 230, output vessel 250 and optional secondary reaction vessel 220, waste vessel 240, and quality control module 260) ) may be formed integrally with the base 202.

In some embodiments, output vessel 250 may include a separate removable component, such as an Eppendorf tube. This may allow the final output liquid to be easily removed from the cartridge 200 within a sealed container 250 for further processing or use elsewhere.

Sample cartridge 200 may define an output container holder or sheet 254 that may be formed integrally with base 202. Output container 250 may be seated on seat 254 during processing in device 100 . Once the selected instrument workflow is complete and the output fluid has been deposited into the output container 250, the output container 250 can be removed from the seal and seat 254 and the remainder of the sample cartridge 200 can be discarded. ) can be.

2G, a flow schematic diagram of a sample cartridge 200 is shown with optional additional features according to some embodiments. The network of channels and valves of sample cartridge 200 will be described with reference to a simple workflow, but it will be understood that many other workflows can be performed on or within cartridge 200.

A liquid sample can be introduced into the primary reaction vessel 210 and a lid 211 is used to seal the sample within the primary reaction vessel 210. The lid 211 may be formed integrally with the primary reaction vessel 210, for example, as shown in FIG. 2A.

One or more reagents may be dispensed into the open top of reagent vessel 230 (e.g., from reagent module 300). The primary reagent channel 231 extends between the reagent vessel 230 and the primary reaction vessel 210. Reagents may be transferred from the reagent vessel 230 to the primary reaction vessel 210 through the primary reagent channel 231.

Primary reagent valve 235 may be disposed in primary reagent channel 231 to control flow through primary reagent channel 231. Primary reagent valve 235 may include an active valve (examples of which are discussed below) or a passive valve. For example, primary reagent valve 235 may include a low pressure valve, which has a relatively low cracking pressure compared to certain other valves in the network. That is, the valve may restrict flow until a relatively low critical pressure difference exists across the valve, at which point the primary reagent valve 235 opens and fluid flows through the primary reagent channel 231 into the reagent vessel. It can flow from 230 to the first reaction vessel 210. .

The driving pressure gradient can be created using the pneumatic module 500. Cartridge 200 may include a primary pneumatic channel 212 extending between the primary reaction vessel 211 and the primary pneumatic port 213. Primary pneumatic port 213 may be defined by openings in the outer surface of sample cartridge 200, such as the lower surface or side surface of base 202, along with other pneumatic ports described below. The device 100 may be configured to engage with pneumatic connectors 510 to connect the pneumatic port 213 to the pneumatic module 500.

The pneumatic module 500 may include a plate defining a plurality of pneumatic ports each connected to a pressure control manifold by a pneumatic line. Each pneumatic port may include a seal and may be configured to connect to a corresponding port on the underside of cartridge base 202. The plate may be configured to be moved upward by a motion module to meet the cartridge once the cartridge is installed in the device, whereby corresponding ports are connected to connect the pneumatic module to the channels of the cartridge so that they are in fluid communication. Let it happen.

With the lid 211 sealed, when the pneumatic module 500 applies negative or vacuum pressure to the pneumatic port 213 (negative relative to atmospheric or ambient pressure), a pressure gradient occurs in the primary reaction vessel 210. and the reagent vessel 230 so that reagents can be drawn from the reagent vessel 230 through the primary reagent channel 231 and flow into the primary reaction vessel and the sample contained therein.

Primary reagent valve 235 remains closed until activated to open or until the critical cracking pressure is overcome by pressure applied to pneumatic port 213 by pneumatic module 500. The flow of the primary reagent channel 231 may be restricted. Primary reagent valve 235 may include a check valve configured to restrict or prevent backflow to prevent a portion of the fluid sample within primary reaction chamber 210 from flowing into reagent vessel 230 .

To avoid a portion of the contents of the primary reaction vessel 210 being drawn into the primary pneumatic channel 212, the opening of the primary pneumatic channel 212 into the primary reaction vessel 210 is shown in Figure 2A. As shown, it may be defined over a portion of the side wall of the primary reaction vessel 210 or at or near the top of the primary reaction vessel 210. The primary pneumatic channel 212 may be defined as a structure extending over the side of the primary reaction vessel 210 within or along the side wall of the primary reaction vessel 210, as shown in FIG. 2A.

In some embodiments, primary reagent channel 231 may also extend upwardly along the sidewall and at or near the top of primary reaction vessel 210 as shown in Figure 2A. It can be opened with This may further reduce the likelihood that a portion of the fluid sample will flow from the primary reaction vessel 210 into the primary reagent channel 231 or reagent vessel 230.

Alternative designs for integrating the pneumatic channel and input channel of one of the reaction vessels 210, 230 into the sample cartridge 200 are shown in FIGS. 2H and 2I. For example, the channels may be formed as open channels in the base 202 and the flat perpendicular web 203 as shown in FIG. 2H. Alternatively, the channels may be formed as open channels in the side structures 204 extending upwardly with the reaction vessels 210, 230 as shown in Figure i. The channels can then be closed by covering them with a foil or film that can be adhesively bonded or welded to the base 202 and web 203 or side structure 204.

If the instrument workflow includes operations that require removal of waste fluid from the primary reaction vessel 210, the sample cartridge 200 may include a waste container 240. Alternatively, device 100 may include a waste receptacle or waste channel for externally disposing waste fluid.

Sample cartridge 200 may include a primary waste channel 214 extending between primary reaction vessel 210 and waste vessel 240 (or other waste channel or bowl). A primary waste valve 215 may be disposed in the primary waste channel 214 to control when fluid is removed from the primary reaction vessel 210 through the primary waste channel 214. For example, primary waste valve 215 may include a low pressure valve with a relatively low cracking pressure.

Sample cartridge 200 may further include a waste pneumatic channel 242 extending between waste container 240 and waste pneumatic port 243. The waste pneumatic channel 242 may also open into the waste container 240 at or near the top of the waste container 240 to avoid drawing waste liquid into the waste pneumatic channel 242. The top of waste container 240 may be sealed, for example with a lid or foil.

In some embodiments, depending on which liquids are to be transferred between the vessels 210, 220, 230, 240, some splashing may occur and a small amount of liquid may flow into the pneumatic channels 212, 222, 242. You can fry it. If liquids are drawn through the pneumatic channels, they can pass from the cartridge to the pneumatic module 500 and contaminate the device.

To alleviate this scenario, the cartridge may include liquid traps associated with one or more (or each) of the pneumatic channels to prevent or restrict liquids leaving the cartridge through the pneumatic channels. For example, liquid traps may include gas permeable membranes that allow passage of air but restrict or stop the passage of liquids. The gas permeable membrane may be positioned anywhere along the pneumatic channels 212, 222, 242, such as at the openings or at the end of each pneumatic channel in the base of the cartridge.

In some embodiments, gas permeable membranes are disposed over a relatively large area (larger than the cross-section of the corresponding channel) to increase the capacity of trapped liquid that can be present before blocking the flow of gas through the membranes. It can be. The device may be configured to detect a change in pressure due to a blockage of one of the channels or liquid traps, which may then trigger, for example, workflow operations to end and indicate that the process has failed.

Sample cartridge 200 may further define a primary output channel 216 to allow discharge of output fluid from primary reaction chamber 210. In some embodiments, when only one reaction vessel is needed, primary output channel 216 may lead directly to output vessel 250. In some embodiments, if a secondary reaction vessel is desired, the primary output channel 216 may extend between the primary reaction vessel 210 and the secondary reaction vessel 220.

A primary outlet valve 217 may be disposed in the primary outlet channel 216 to control discharge of output fluid through the primary output channel 216. For example, primary outlet valve 217 may include an active pressure actuated valve that is operated by applying pressure to a corresponding primary outlet valve pneumatic port 218.

In embodiments where the sample cartridge 200 includes a secondary reaction vessel 220, the sample cartridge 200 has a secondary reagent channel 232 extending between the reagent vessel 230 and the secondary reaction vessel 220. ) may include.

Secondary reagent valve 236 may be disposed in secondary reagent channel 232 to control the flow of reagents through secondary reagent channel 232. For example, secondary reagent valve 236 may include a high pressure valve with a relatively high cracking pressure compared to other valves in sample cartridge 200.

Sample cartridge 200 may include a secondary pneumatic channel 222 extending between the secondary reaction vessel 220 and the secondary pneumatic port 223. The secondary pneumatic channel 222 may open into the secondary reaction vessel 220 at or near the top of the secondary reaction vessel 220. The top of waste container 240 may be sealed, for example with a lid or foil.

Negative vacuum pressure is applied to the secondary pneumatic port 223 to pull the reagents from the reagent vessel 230 into the secondary reaction vessel 220 through the secondary reagent channel 232. This can create a pressure difference sufficient to overcome the relatively high cracking pressure across the secondary reagent valve 236. During this flow, primary output valve 217 may be closed to avoid flow through primary outlet output 216.

Meanwhile, when the flow of output fluid is required from the primary reaction vessel 210 to the secondary reaction vessel 220, the primary output valve 217 can be opened and vacuum pressure is applied to the secondary pneumatic port 223. It can be applied to create a pressure difference sufficient to induce flow through the primary output channel 216 but not sufficient to overcome the relatively high cracking pressure of the secondary reagent valve 236. The primary output channel 216 may open into the secondary reaction vessel 220 at or near the top of the secondary reaction vessel 220.

Sample cartridge 200 may include a secondary waste channel 224 extending between secondary reaction vessel 220 and waste vessel 240 (or other waste channel or vessel). A secondary waste valve 225 may be disposed in the secondary waste channel 224 to control when fluid is removed from the secondary reaction vessel 220 through the secondary waste channel 224. For example, secondary waste valve 225 may include a low pressure valve with a relatively low cracking pressure. In some embodiments, secondary waste channel 224 may not be needed. That is, this is the case where there is no waste liquid to be removed from the secondary reaction vessel 220.

Sample cartridge 200 may further define a secondary output channel 226 to allow discharge of output fluid from secondary reaction chamber 220. In some embodiments, if quality control is not required, secondary output channel 226 may be connected directly to output vessel 250. In some embodiments, if quality control is required, secondary output channel 226 may be connected directly to output vessel 250. ) can extend between the secondary reaction vessel 220 and the quality control module 260.

Secondary output channel 226 may extend between secondary reaction vessel 220 and buffer junction 228. A secondary outlet valve 227 may be disposed in the secondary output channel 226 to control the discharge of output fluid through the secondary output channel 226. For example, secondary outlet valve 227 may include a high pressure valve with a relatively high cracking pressure.

Quality control (QC) module 260 includes a quality control QC vessel configured to receive a volume of output fluid from secondary reaction vessel 220 (or primary reaction vessel 210 if no secondary vessel is present) for analysis. 261). Sample cartridge 200 may further include a QC pneumatic channel 262 and a QC pneumatic port 263, wherein vacuum pressure can pull the output fluid from the secondary output channel 226 to the QC vessel 261. It can be applied so that The top of QC container 261 may be sealed, for example with a lid or foil.

In some embodiments, QC vessel 261 may be preloaded with dye (optionally dried dye) to facilitate optical analysis with optical module 400.

In some embodiments the output fluid may be mixed with a quality control buffer prior to optical analysis. The QC buffer may be stored in the QC buffer container 265 before being transferred to the QC container 261 with the output fluid. For example, QC buffer vessel 265 may define an open top so that buffer can be dispensed into QC buffer vessel 265 by reagent module 300.

Sample cartridge 200 has a QC buffer channel 266 extending from QC buffer vessel 261 at buffer junction 228 to secondary output channel 226 (or primary output channel 216 if there is no secondary vessel). ) may include. A QC buffer valve 267 may be disposed in the QC buffer channel 266 to control the flow of buffer through the QC buffer channel 266. For example, QC buffer valve 267 may include an active valve, such as a pressure-actuated valve, that is activated by applying positive or negative pressure to a corresponding QC buffer pneumatic port 268.

Sample cartridge 200 further includes a metering channel 299 in fluid communication with secondary output channel 226 and buffer channel 266 and extending from buffer junction 228 to quality control junction 229. can do.

Sample cartridge 200 may further include a QC channel 269 extending between QC junction 229 and QC vessel 261. The sample cartridge 200 may further include a QC vessel valve 264 disposed in the QC channel 269 to control the flow of fluid through the QC channel 269 into the QC vessel 261. QC vessel valve 264 may include a low pressure valve having a relatively low cracking pressure.

With the QC buffer valve 267 closed, a portion of the output fluid flows from the secondary reaction vessel 220 through the buffer junction 228 into the secondary output channel 226 and into the metering channel (229) to the QC junction 229. Vacuum pressure may be applied to the QC vessel pneumatic port 263 to create a relatively high pressure differential to overcome the threshold of the secondary output valve 227 for a short period of time to pull the pressure differential to 299). can be neutralized and stop the flow. Metering channel 299 may define an accurate aliquot of output fluid by defining a known volume (e.g., 1 μL) such that metering channel 299 can be filled from buffer junction 228 to QC junction 229. .

The QC buffer valve 267 then applies an appropriate activation pressure to the QC buffer pneumatic port 268 and a vacuum pressure to the QC vessel pneumatic port 263, sufficiently high to open the low pressure QC vessel valve 264, but high enough to open the high pressure QC vessel valve 264. It can be opened by creating a pressure difference below the threshold for the auxiliary output valve 227. This allows QC buffer to flow through QC buffer channel 266 and through metering channel 299 and QC channel 269 into QC vessel 261 along with an aliquot of output fluid from metering channel 299. .

The mixed fluid can then be mixed with the pre-loaded dye in QC vessel 261 for analysis.

In some embodiments, the sample cartridge 200 may further include one or more QC reference vessels 271, each having a corresponding QC reference pneumatic channel 272 and a QC reference pneumatic port 273; Each of them can be preloaded with a predefined amount of dried dye. Each QC reference vessel 271 also has a corresponding QC buffer vessel 275 configured to receive a specific required amount of QC buffer to be drawn into the QC reference vessel 271 by applying vacuum pressure to the corresponding QC reference pneumatic port 273. You can have it.

The contents of the QC vessel 261 may be compared to the contents of the QC reference vessel 271 using the optical module 400 to measure properties of the output fluid, such as, for example, the concentration of specific components.

Sample cartridge 200 branches off from secondary output channel 226 at QC junction 229 and adds a final output channel 256 connecting secondary output channel 226 to output vessel 250. Included as. A final output valve 257 is disposed in the final output channel 256 to control flow through the final output channel 256. The final output valve 257 may comprise, for example, a low pressure check valve.

Sample cartridge 200 further includes an output vessel pneumatic channel 252 extending between output vessel 250 and output vessel pneumatic port 253. Vacuum pressure may be applied to the output vessel pneumatic port 253 to pull output fluid into the output vessel 250 through the final output channel 256.

The final output channel 256 and output vessel pneumatic channel 252 may be connected to a temporary removable lid 259 (shown in FIGS. 2F and 2G) used to seal the output vessel 250 closed during instrument workflow. You can. Once the sample has been processed and the sample cartridge 200 is removed from the instrument 100, the temporary lid 259 can be removed from the output container 250 and the output container 250 can be closed with the primary output container lid 251. You can. For example, the output container lid 251 may be a hinged lid formed integrally with the output container 250 as shown in FIG. 2A.

To measure an accurate aliquot of output fluid for QC analysis, the vacuum pressure is increased when the output fluid fills the metering channel 299 between the QC junction 229 and the buffer junction 228 and enters the final output channel 256. It can be applied for a predetermined period of time up to. The length of the measurement channel 299 can be designed to define a specific, known volume (e.g., 1 μL). Flow through the final output channel 256 can be stopped by returning the pressure at the output vessel pneumatic port 253 to ambient pressure.

The QC buffer valve 267 can then be opened and vacuum pressure is applied to the QC pneumatic port 263 to pull the buffer from the QC buffer vessel 265 past the QC and buffer junctions 228, 229 and into the metering channel. An aliquot of output fluid is conveyed through 299 and QC channel 269 and drawn into QC vessel 261. In this way an accurate aliquot is defined between the QC and buffer junctions 228, 229, which is slugged into the QC vessel to be mixed with the buffer.

The entire contents of QC buffer vessel 265 may be drawn into QC vessel 261 to ensure that no buffer remains in the channel between the QC and buffer junctions 228, 229. This ensures that a known volume (or very close to a known volume) of buffer is drawn into QC vessel 261. Additionally, it can be advantageous when a high concentration of output fluid is required because it reduces or minimizes the amount of buffer that remains in the channel and can dilute the output fluid.

In some embodiments, sample cartridge 200 may include a waste channel 279 to dispose of excess fluid into a waste container 240 as shown in FIG. 2G. However, in some embodiments, this may not be necessary because the exact volumes of output fluid and buffer are measured and drawn into QC vessel 261 to ensure there is no excess fluid.

In some embodiments, sample cartridge 200 may further include an intermediate outlet 280 from final output channel 256 between final output valve 257 and output vessel 250. Outlet 280 may be sealed with an air-permeable membrane 281 that is permeable to gases but impermeable to liquids. On the other side of membrane 281 (opposite channel 256), a mid-outlet pneumatic channel 282 connects outlet 280 to mid-outlet pneumatic port 283. Air may be drawn through the breathable membrane 281 by applying vacuum pressure to the middle outlet pneumatic port 283.

When this is done, the liquid output fluid will be drawn along the final output channel 256 but will stop once it reaches the breathable membrane 281. This will result in an increase in pressure gradient that can be detected by pneumatic module 500 indicating that channel 256 has filled to intermediate outlet 280. This signal can be used to trigger the next step in the workflow, such as flowing buffer through metering channel 299 and QC channel 269.

Sealed reaction vessels (210, 220), QC vessel (261), QC reference vessel (271), output vessel (250) and waste vessel (240) are subject to cross-section due to splashing of fluid sample out of these vessels. Allows fluid samples to be processed without contamination or instrument contamination. This provides separate containers for receiving reagents in the reagent module (e.g., reagent vessel 230 and buffer vessels 265, 275) and seals the reagents for processing using the pneumatic module and corresponding pneumatic ports. This is achieved by creating pressure gradients that induce flow as needed by transferring the pressure to the vessels. Locating the inlet channels and openings of the pneumatic channels at or near the top of the sealed containers reduces the possibility of sample fluid flowing back into the inlet channels or being drawn into the pneumatic module and contaminating the instrument.

Referring to Figure 2E, a bottom view of sample cartridge 200 is shown illustrating the cartridge's network of channels and valves in more detail. An enlarged view of quality control module 260 is shown in Figure 2F. Similar elements are indicated by like reference numerals.

In some embodiments, QC module 260 may be included as part of a larger sample cartridge, such as sample cartridge 200 or any other sample cartridge that may require QC analysis or accurate fluid metering. In some embodiments, QC module 260 may be included as part of a measurement or analysis instrument.

Referring to Figure 2F, QC module 260 may also be considered independently as a separate sample cartridge 290 according to some embodiments. Some embodiments relate to an independent sample cartridge 290 that includes only a QC module 260, as shown in FIG. 2F.

Sample cartridge 290 may be configured for use with a fluid analysis instrument, for example. Cartridge 290 includes a sample vessel 220 configured to receive a fluid sample for analysis. The sample vessel 220 may correspond to the secondary reaction vessel 220 of the sample cartridge 200, or may correspond to the primary reaction vessel 210 of the sample cartridge 200 without the secondary reaction vessel 220.

Cartridge 290 includes a buffer container 265 (similar to sample cartridge 200) configured to contain buffer. Cartridge 290 includes a sealed assay vessel 261 (corresponding to QC vessel 261) configured to receive a mixed fluid comprising an aliquot of a fluid sample mixed with at least a portion of a buffer for analysis.

Cartridge 290 includes a sample channel 226 (corresponding to secondary output channel 226) extending between sample vessel 220 and first junction 228 (corresponding to buffer junction 228).

Cartridge 290 includes a sample channel valve 227 (corresponding to secondary output valve 227) disposed in sample channel 226 to control the flow of sample through sample channel 226.

Cartridge 290 includes a buffer channel 266 extending between buffer container 265 and first junction 288. A buffer channel valve 267 is disposed in the buffer channel 266 to control the flow of buffer solution through the buffer channel 266.

Cartridge 290 includes a metering channel 299 in fluid communication with a buffer channel 266 and a sample channel 226, wherein the metering channel 299 has a first junction 228 and a second junction 229 (QC (corresponding to the joint 229) extends between them.

Cartridge 290 is in fluid communication with metering channel 299 and includes an assay vessel channel 269 (corresponding to QC channel 269) extending between second junction 229 and assay vessel 261. The cartridge 290 includes an analytical vessel pneumatic port 263 (corresponding to the QC pneumatic port 263) in communication with the analytical vessel 261 and is connected to a pneumatic module to selectively adjust the pressure of the analytical vessel 261 to control the analytical vessel 261. It is configured to draw fluid through channel 269 into analysis vessel 261.

At least one of the sample channel valve 227 and the buffer channel valve 267 may include an active valve that can be selectively opened and closed to draw an aliquot of the fluid sample into the metering channel 299, wherein the buffer Allows to be drawn through buffer channel 266 and through metering channel 299 and analysis vessel channel 269 into analysis vessel 261 along with an aliquot of fluid sample for analysis. For example, buffer channel valve 267 may include an active valve and sample channel valve 227 may include a relatively high pressure check-valve.

Assay vessel 261 may be preloaded with a dye configured to mix with the buffer and fluid sample to facilitate analysis.

Sample cartridge 290 or 200 may further include an intermediate outlet 280 in fluid communication with metering channel 299 through second junction 229. Middle outlet 280 may be similar to the middle outlet of sample cartridge 200, and both may include one of the features described below.

Referring to Figure 2O (adjacent Figure 2F), an enlarged top perspective view of outlet 280 according to some embodiments is shown.

In some embodiments, outlet 280 may be located at second junction 229. In some embodiments, outlet 280 can be located distal to second junction 229 and connected to second junction 229 via outlet channel 285.

Sample cartridge 290 or 200 may include an outlet chamber 284 with an intermediate outlet 280 opening. A breathable liquid barrier membrane 281 may cover the outlet 280.

A sample cartridge (290 or 200) is in fluid communication with the outlet chamber (284) and includes a pneumatic module to selectively adjust the pressure in the outlet chamber (284) to draw air from the metering channel (299) through the breathable membrane (281). It may include a middle outlet pneumatic port 283 configured to be connected to.

Sample cartridge 290 or 200 may include an intermediate outlet pneumatic channel 282 extending between intermediate outlet pneumatic port 283 and outlet chamber 284. Outlet chamber 284 can be sealed with only two fluid openings: outlet 280 and the opening of the intermediate outlet pneumatic channel 282. The top of outlet chamber 284 may be sealed, for example with foil.

The intermediate outlet 280 allows liquid drawn from the sample channel 226 or buffer channel 266 into the metering channel 299 to fill the metering channel 299 but not to proceed into the analysis vessel channel 269. It can be arranged so that

The sample cartridge 290 or 200 may allow liquid to be drawn from the sample channel 226 into the metering channel 299, or the buffer channel 266 may fill the metering channel 299 and proceed to the outlet channel 285, but may also proceed to the outlet channel 285. It may include an outlet channel 285 extending between the second junction 229 and the outlet 280 such that it is not permitted to proceed to 269 .

Sample cartridge 290 or 200 may include an output vessel 250 in fluid communication with metering channel 299 through second junction 229 and output channel 256. Output channel 256 may extend from second junction 229 to output container 250. Alternatively or additionally, the output channel may extend between the intermediate outlet 280 and the output container 250.

The sample cartridge 290 or 200 communicates with the output vessel 250 and is configured to draw fluid from the metering channel 299 to the output vessel 250 through the second junction 229 and the output channel 256. 250) may include an output vessel pneumatic port 253 configured to be connected to a pneumatic module to selectively adjust the pressure of the vessel.

The arrangement of channels, valves, vessels and outlets in sample cartridge 290 and QC module 260 accurately quantifies aliquots of sample fluid (or treated fluid) because the volume of the metering channel can be precisely defined. make it possible The arrangements described above can be used to accurately meter fluids for quantitation in any application where accurate quantification is required.

Sample cartridge 290 further includes one or more reference vessels 271 and associated buffer vessels 275, pneumatic channels 272, and pneumatic ports 273, as described with respect to sample cartridge 200. It can be included.

Output channel 256 and output vessel pneumatic channel 252 may be connected to a temporary lid 259 that defines openings of output channel 256 and output vessel pneumatic channel 252 into output vessel 250, It may be configured to seal (250). The temporary lid 259 can be removed so that the output container 250 can be removed from the base 202 of the sample cartridge 290, and the output container 250 can be sealed with the output container lid 251.

Sample cartridge 200 or 290 may be formed from any plastic material appropriate for a given application. For example, polypropylene can be used to handle biological materials.

Sample cartridge 200 or 290 may be formed by injection molding. For example, sample cartridge 200 or 290 may be formed of some or all of channels, chambers and vessels with an open top or open side, some of which have openings, for example sealed as described above. The channels, chambers and vessels can be sealed with welded foil as needed to form them.

The channels in the base can be covered with a polypropylene membrane that can be heat welded to the base. And the channels in the side walls connecting the vessels can similarly be covered with a polypropylene membrane heat welded to the body. For example, heat welding may include laser welding, and tractor welding around the perimeter of each channel may secure the membrane to the body to define each channel.

The valves may include any suitable active or passive valves depending on the arrangement and application. Suitable valves include check valves with different relative cracking pressures (e.g., as shown in Figure 2J), duck-bill valves (e.g., as shown in Figure 2L), umbrellas, etc. (umbrella) valves (e.g., as shown in Figure 2M), microfluidic valves or capillary valves with different cracking pressures depending on surface tension and capillary action, Pressure actuated switch valves (e.g., as shown in Figure 2K), electronically actuated switch valves (e.g., solenoid valves) or Prodger valves (e.g., as shown in Figure 2n) (as described above) may include mechanically operated switch valves. A combination of different valve types may be used to achieve the required flows in sample cartridge 200.

3A and 3B, a reagent module 300 is shown according to some embodiments. The reagent module 300 includes a plurality of reagent cartridges 320 removably mounted on the frame 350. Frame 350 also supports a pump 360 configured to control the dispensing of reagents in reagent cartridges 320.

An isolated reagent cartridge 320 is shown in FIG. 3B. Each reagent cartridge 320 is configured to support a reservoir 322, a flexible distribution tube 323, and the reservoir 322, and engages the reagent module frame 350 to hold the reagent cartridge 320 in the frame 350. ) includes a reagent cartridge frame 325 configured to be mounted on the

Pump 360 may include a peristaltic pump. For example, the pump 360 may include a pump shaft 363 and one or more motors 362 each configured to drive rotation of a pump cam (not shown) mounted on the pump shaft 363. The pump cam may generally be circular with protuberances, for example round toothed cogs.

When mounted on the reagent module frame 350, the reagent cartridge frame 325 supports the distribution tube 323 such that a portion of the tube 323 extends at least partially around the circumference of the pump cam and the corresponding motor ( When the pump cam is rotated by 362), the protrusions of the pump cam contact and compress a portion of the distribution tube 323, thereby pushing fluid through the distribution tube 323 as the pump cam rotates. The dispensing tube 323 can be positioned such that when the reagent module 300 is aligned with the sample cartridge 200, the openings dispense reagents into a desired vessel (reagent vessel or QC buffer vessels) of the sample cartridge 200.

Distribution tube 323 may be formed of any suitable material, and in some cases may be formed of other materials depending on compatibility with individual reagents. For example, distribution tube 323 may be formed from silicone, viton, or Chem-Durance Bio tubing.

Each reagent cartridge 320 may be engaged by a separate pump cam driven by a corresponding one of the motors 362. In some embodiments a single pump shaft 363 configured to drive rotation of a single pump cam or multiple pump cams may be configured to engage one or more reagent cartridges 320 to control dispensing. For example, if multiple reagents need to be dispensed simultaneously in similar amounts, corresponding reagent cartridges 320 can be simultaneously engaged by a single pump system to dispense reagents into sample cartridge 200.

Pump 360 may include independent motors 362 and drive shafts 363 to independently control the dispensing of reagents from different reagent cartridges 320 .

Reservoirs 322 of different reagent cartridges 320 may define different volumes proportional to the expected rate of consumption of the different reagents contained therein. For example, if the first reagent is typically dispensed in twice the volume of the second reagent, the first reagent reservoir 322 may be twice the volume of the reservoir 322 for the second reagent.

Reservoirs 322 may contain sufficient quantities of reagents for a certain number of instrument workflows to be performed. When the reagent reservoirs 322 are empty, the reagent module 300 can be partially slid out of the instrument housing 110 as shown in FIG. 1B so that the reagent reservoirs 322 can be refilled, or The empty reagent cartridge 320 can be completely removed and replaced with a filled reagent cartridge 320.

In some embodiments, the reagent module frame 350 may be mounted on or include the carriage 920 of the motion module 900. Reagent module 300 moves axis 903 across a plurality of sample cartridges 200 in cartridge slots 120 to dispense reagents to sample cartridges 200 at selected times during instrument workflows. ) can be moved (by the motion module 900) along.

The reagent module 300 may be moved to a selected carriage position at a selected time corresponding to the selected sample cartridge 200. Pump 360 may then be operated to dispense one or more reagents from corresponding reagent cartridges 320 to a selected container of sample cartridge 200, such as, for example, reagent vessel 230 or quality control buffer vessels. there is.

4A and 4B, a diagram of an optical module 400 according to some embodiments is shown. The optical module 400 includes a light source 410 and a detector 420. Light source 410 is configured to illuminate a quality control (QC) sample 404 , which may include the output liquid of QC vessel 261 or a reference liquid from one of the QC reference vessels 271 . Detector 420 is configured to detect and measure light transmitted from QC sample 404.

For example, the light source 410 may include an LED or a laser. Detector 420 may include a photodiode or any other suitable optical detector. Light source 410 and detector 410 may operate in any suitable frequency range, including visible, near visible, infrared, and ultraviolet ranges, depending on the application and the property being measured. It can be configured to operate.

The optical module 400 may be configured to measure any one or more of scattered light, refracted light, or reflected light transmitted from the QC sample 404.

In some embodiments, optical module 400 may include one or more lenses, filters and/or other optical devices. For example, optical module 400 may include a source lens 412 for focusing light from source 410 (e.g., into collimated beams); a beam splitter 414 to redirect light from source 410 toward QC sample 404; a sample lens 402 for focusing the source light onto the QC sample 404 and refocusing the light transmitted from the QC sample 404 (e.g., into collimated beams); a detector lens 422 for focusing light transmitted from the QC sample 404 onto a detector 420; and one or more filters 430 disposed in the detector path and/or source path to filter specific frequencies of light.

The optical module 400 is mounted on the carriage 920 of the motion module 900 (the same carriage as the reagent module 300 or an independent carriage 920) so that the optical module 400 is placed on the sample cartridge 200. The QC sample 404 may be allowed to be aligned with a selected one sample cartridge 200 in the cartridge slots 120 for optical analysis.

For example, the sample cartridge 200 shown in Figure 2A includes one QC vessel 261 and three QC reference vessels 271 to be compared. The QC vessel 261 and the three QC reference vessels 271 are arranged along the lateral axis of the sample cartridge 200 parallel to the movement axis 904 of the optical module 404 to This allows easy access to QC samples 404. The carriage 920 can be moved by the motion module 900 to different carriage positions corresponding to the positions of the QC vessel 261 and the three QC reference vessels 271 .

QC vessel 261 and three QC reference vessels 271 direct light from the optical source 410 to the QC samples 404 contained therein and from the QC samples 404 to the optical detector 420. It may contain a transparent window (top, bottom or side) that allows passage. Windows may have a surface finish rated SPI A-1 to minimize scattering. The window thickness may be less than or equal to 3 mm.

Motion module 900 may be positioned with optical module 400 such that QC sample 404 is within 2 mm of the optical focal plane of optical module 400, and optionally within a lateral position tolerance of 1.25 mm. Different tolerances may be appropriate for different applications depending on the characteristics of the optical module 400 and sample cartridge 200.

A pneumatic module 500 according to some embodiments is shown in FIG. 1C. The pneumatic module 500 may include a pressure regulator, a compressor, a vacuum generator, or a vacuum pump. Alternatively, pneumatic module 500 may be configured to be connected to an external pressure source or vacuum line, for example.

Pneumatic module 500 may include a network of pneumatic lines and valves with connectors adjacent cartridge slots 120 configured to connect to pneumatic ports of sample cartridges 200 . Pneumatic module 500 may be configured to selectively deliver positive and/or negative pressure (relative to atmospheric pressure) to selected pneumatic ports of sample cartridges 200 at selected times during instrument workflows.

In some embodiments, pneumatic module 500 may be configured to deliver different magnitudes of relative pressure differences to different pneumatic ports of sample cartridges 200. In some embodiments, pneumatic module 500 may be configured to deliver different magnitudes of relative pressure differences to selected pneumatic ports of sample cartridges 200 at different times. In some embodiments, the pneumatic module 500 may be configured to deliver negative pressure only to the pneumatic ports of the sample cartridges 200.

In some embodiments, pneumatic module 500 may include sets of pneumatic lines, each set of pneumatic lines applying a selected pressure to all of the plurality of sample cartridges 200 in cartridge slots 120. It is configured to deliver simultaneously to corresponding pneumatic ports. For example, a constant negative pressure is applied to all primary pneumatic ports 213 at the same time.

In some embodiments, each pneumatic port of sample cartridge 200 may have a single selected pressure or pressure range to be delivered at selected times without the need to change the pressure.

Pneumatic module 500 may include any suitable valve system to selectively deliver the required pressure to the pneumatic ports required at selected times of instrument workflows. For example, an array of solenoid valves electronically actuated by control module 101, or a manifold valve system or a rotary valve system.

5A and 5B, components of a pneumatic module 500, a thermal module 600, a magnetic module 700, a mixing module 800, and a motion module 900 according to some embodiments are shown, and the chassis Alternatively, they may be received within housing 110 and cooperate to form a core unit 1100 that defines a cartridge slot or socket 120 .

Pneumatic connectors 510 are shown below cartridge slots 120 supported by pneumatic support frame 505 . The support frame 505 is connected to a pneumatic module actuator 905 , which may form part of the motion module 900 . Pneumatic module actuator 905 may include a motor or linear actuator configured to raise and lower pneumatic connectors 510 . Pneumatic connectors 510 may be in a lowered position for loading (or removing) sample cartridge 200 from cartridge slots 120 . When the sample cartridges 200 are received within the cartridge slots 120, the pneumatic module actuator 905 causes the pneumatic connectors 510 to engage pneumatic ports within the sample cartridges 200 and actuate the pneumatic module 500. Can be operated to raise the pneumatic support frame 505 and the pneumatic connectors 510 to fluidly connect to the channels of the sample cartridge 200.

In some embodiments, motion module 900 may further include a vertical movement platform 950 configured to raise and lower other components of device 100 within housing 110 . Movement of the platform 950 is achieved by a platform actuator 955, which may include, for example, a motor 957 and a linear actuator with a lead screw 959 or a leadscrew actuator as shown in FIG. 4B. It can be driven by

Vertical movement platform 950 may be configured to raise and lower components of, for example, thermal module 600 and/or magnetic module 700 as well as any other components of the appliance that may require raising and lowering. there is. In some embodiments, pneumatic support frame 505 and connectors 510 may be mounted on a similar vertically moving platform 950. In some embodiments, motion module 900 may include multiple vertical movement platforms 950 configured to operate independently to independently raise and lower different components or groups of components. For example, the thermal module and the magnetic module may be mounted on a single vertically movable platform, which may include an additional vertically movable platform mounted thereon to raise and lower the thermal module independently of the magnetic module.

In some embodiments, device 100 may not include vertically moving platform 950 .

Thermal module 600 may include one or more thermal control devices 610, including heating and/or cooling elements, thermoelectric devices, peltier elements, resistive heaters, heat lamps, thermal It may include exchangers and/or fans. When heating (or other thermal control) is required during one of the instrument workflows (e.g., for culturing), the platform 950 places thermal control devices 610 (e.g., heaters) in the cartridge slot. can be brought closer to the sample cartridges 200 and raised to activate the thermal control devices 610 .

In some embodiments, thermal module 600 may include a conductive member coupled to a heating element to facilitate heating of reaction vessels. For example, the conductive member can include a plate or jacket that can define a complementary surface configured to partially surround the reaction vessel.

Thermal module 600 may include a cooling fan disposed beneath the heating elements and conductive members and configured to direct ambient air upward around the heating elements and conductive members to cool them when needed.

The cartridge is configured to allow the passage of conductive elements or magnets and/or magnets for positioning next to the reaction vessels 210, 220 of the primary reaction vessel 210 and/or secondary reaction vessel 220. Openings may be defined in the base 202 around the bottom.

In some embodiments, thermal module 600 is secured in a position proximate to cartridge slots 120 and aligned with primary and/or secondary reaction vessels 210, 220, depending on the thermal adjustments required for the particular workflow operation. Depending on the temperature, it can simply switch from heating to cooling or neutral.

The magnetic module 700 may include one or more magnets 710 arranged to control the movement of magnetic beads in the primary and/or secondary reaction vessels 210, 220. Magnets 710 may include permanent magnets and/or electromagnets and are mounted on a vertically moving platform 950 to allow the magnetic beads to react in the primary and/or secondary reaction vessels 210, 220 when held stationary. ) and the magnets 710 may be lowered further away from the primary and/or secondary reaction vessels 210, 220 to have less impact on the magnetic beads. Magnets 710 are placed on either side of the primary and/or secondary reaction vessel 210, 220 to position the beads away from the outlet outlets to prevent blockage or constriction of the outlet flow into the channels. maintain them.

In some embodiments, for example, when magnets 710 include electromagnets, magnets 710 are secured in a position proximate to cartridge slots 120 and primary and/or secondary reaction vessels 210, 220, and can be simply turned on or off.

In some embodiments, the device may not include a magnetic module 700 and the functionalized beads in the primary and/or secondary reaction vessels may be kept outside the output channels, for example by a physical barrier or restriction, such as a filter. It can be maintained.

Mixing module 800 may include any suitable device to enhance mixing of fluids in sample cartridges 200. For example, mixing module 800 may include a shaker 810. Shaker 810 may include an orbital shaker, such as an eccentric cam or offset weight configured to be rotated by a motor to induce vibrations in device 100. Alternatively, other conventional mixing devices may be used to facilitate mixing of fluids in sample cartridge 200. In some embodiments, mixing module 800 includes a single shaker 810 configured to shake all sample cartridges 200 simultaneously.

The orbital shaker can include a number of weights and counter weights configured to vibrate the cartridges without toppling the device. The power, frequency and amplitude of vibration appropriate to the required application can be selected. When processing relatively fragile molecules (e.g., nucleic acids), frequencies below 2000 rpm, for example about 1100 rpm, may be appropriate.

6, a schematic diagram of a control module 101 according to some embodiments is shown. The control module 101 controls the reagent module 300, optical module 400, pneumatic module 500, thermal module 600, magnetic module 700, mixing module 800, and motion module 900. It includes electronic devices and software configured to control operations performed by the device 100 .

Control module 101 may also include one or more sensors to monitor the operations of the modules. Sensors may include, for example, position sensors, accelerometers, proximity sensors, angle sensors (eg, shaft angle or speed sensor), hall sensors, and pressure sensors.

5C-5F, core unit 1100 is shown in greater detail, according to some embodiments, as sample cartridge 200 (or sample cartridge 1000 shown in FIG. 7A), as further described below. )) can be configured to accommodate.

Core unit 1100 may at least partially define a cartridge slot or socket 120 each configured to receive a cartridge 1000. Each cartridge socket 120 may have an associated pneumatic interface plate 1500 (FIG. 5D) configured to engage and connect the cartridge 1000 to the pneumatic module 500 via pneumatic lines 515.

Pneumatic plate 1500 may define pneumatic plate ports 1501, 1502, 1503, 1504, 1505, 1506, 1507, 1508, 1509, and 1510 in fluid communication with pneumatic line 515. The pneumatic plate ports are in turn configured to connect with the corresponding cartridge pneumatic ports 1201, 1202, 1203, 1204, 1205, 1206, 1207, 1208, 1209, 1210 of the cartridge 1000 (FIG. 7B). The pneumatic plate ports may be arranged differently or in different numbers to suit different sample cartridges. The arrangement and connection between the cartridge pneumatic ports and the corresponding pneumatic plate ports on the underside of the sample cartridge 1000 can be understood by comparing Figures 5D and 7B:

Cartridge port 1201 is configured to connect to pneumatic plate port 1501;

Cartridge port 1202 is configured to connect to pneumatic plate port 1502;

Cartridge port 1203 is configured to connect to pneumatic plate port 1503;

Cartridge port 1204 is configured to connect to pneumatic plate port 1504;

Cartridge port 1205 is configured to connect to pneumatic plate port 1505;

Cartridge port 1206 is configured to connect to pneumatic plate port 1506;

Cartridge port 1207 is configured to connect to pneumatic plate port 1507;

Cartridge port 1208 is configured to connect to pneumatic plate port 1508;

Cartridge port 1209 is configured to connect to pneumatic plate port 1509;

Cartridge port 1210 is configured to connect to pneumatic plate port 1510.

The pneumatic module 500 may be configured to independently and selectively adjust the pressure of each pneumatic line 515 to direct liquid flow within the channels of the cartridge 1000 and/or operate the valves of the cartridge 1000. there is.

In some embodiments, the sample cartridge may be configured for positive operating pressure (higher than ambient pressure) in some or all of the pneumatic lines 515. In the illustrated embodiment, sample cartridge 1000 is configured for negative operating pressure (less than ambient pressure) in all pneumatic lines 515.

In some embodiments, the sample cartridge may be configured for operating pressure at a single pressure level. In the illustrated embodiment, sample cartridge 1000 is configured for two operating pressure levels. The first operating pressure may be a relatively high magnitude sound pressure, for example in the range of 180 mBar to 500 mBar, 190 mBar to 350 mBar or about 200 mBar. The second operating pressure may be a relatively low level of sound pressure, for example in the range of 50 mBar to 200 mBar, 80 mBar to 150 mBar, 100 mBar to 120 mBar or about 120 mBar. The difference between the two pressure levels may for example range from 20 mBar to 200 mBar, from 50 mBar to 100 mBar, at least 20 mBar, at least 50 mBar or at least 100 mBar. Cartridge pneumatic ports 1201, 1202, 1203, 1204, 1205 can be configured for a relatively high first operating pressure. Cartridge pneumatic ports 1206, 1207, 1208, 1209, 1210 can be configured for a relatively low second operating pressure.

Socket 120 may include parallel rails 1120 defining grooves configured to slidably receive edges 1220 of cartridge base 202 when inserted into socket 120. Rails 1120 may include recesses 1122 configured to receive elastic clips 1222 formed integrally with cartridge base 202 (FIG. 7F).

Pneumatic interface plate 1500 is positioned below socket 120 and may be biased into the engaged position by springs. The core unit 1100 may include a core carriage 1190 configured to move a pair of lead screws 1191 up and down operated by motors 1192 forming part of the motion module 900. there is. Core carriage 1190 can be seen in Figure 5E with some components of core unit 1100 omitted to better visualize the internal components.

The pneumatic plates 1500 may be connected to the retraction rods 1520 so as to slide up and down the retraction rods 1520 through the core carriage 1190, and the lower ends of the retraction rods 1520 are attached to the core carriage ( 1190) and includes stops 1522 located below. When the core carriage 1190 is lowered beyond engagement with the stops 1522, the retraction rods 1520 and pneumatic interface plates 1500 are lowered with the core carriage 1190.

Retraction of the interface plates 1500 allows the cartridges 1000 to be inserted into the sockets 120 . The operation module 900 raises the core carriage 1190 so that springs raise the interface plate 1500 and press it against the base 202 of the cartridge 1000 to press the interface plate 1500 and the rails ( 1120) can be operated to clamp between them. Pneumatic interface plate 1500 may include gaskets or sealing portions 1530 surrounding each of the pneumatic plate ports 1501, 1502, 1503, 1504, 1505, 1506, 1507, 1508, 1509, 1510. and is configured to be compressed and deformed between the pneumatic plate 1500 and the cartridge 1000 to provide a seal around the connection between the corresponding pneumatic ports 1501 and 1201. Gaskets 1530 may be formed of any suitable elastomeric material, such as rubber, silicone, or other polymers, for example. Gaskets 1530 may define, for example, a truncated cone shape or any other suitable shape to provide a seal between the opposing flat surfaces of pneumatic interface plate 1500 and the bottom of cartridge 1000.

The thermal module 600 includes a radiator 662 at one end and a cooling fan at the opposite end, which may be mounted on a core carriage 1190 or an independent motion stage. It may include heaters 610 including a (cooling fan) 663 (FIG. 5F).

Radiator 662 and magnets 710, as shown in FIGS. 5D and 5E, allow the cartridges to extend through holes 1230 in the base of cartridge 1000, as shown in FIGS. 7B and 5F. Allowing for a slit to be defined to bring the radiator 662, magnets 710, and reaction vessels 210, 220 closer together.

7A-7Z, sample cartridge 1000 is shown in greater detail, according to some embodiments. Sample cartridge 1000 may include similar features to those described with respect to sample cartridge 200, with similar features indicated by like reference numerals.

Referring to Figure 7C, sample cartridge 1000 is shown in an exploded perspective view illustrating the various components that combine to form sample cartridge 1000.

Sample cartridge 1000 includes a body 1001 that defines a base 202, a primary reaction vessel 210, a secondary reaction vessel 220, a reagent vessel 230, and a waste vessel 240. Body 1001 also defines other features described in detail below.

Body 1001 may be formed by any suitable means of any material suitable for the particular application. Some applications may require non-reactive materials suitable for the samples and reagents to be processed. Body 1001 may be injection molded and defined from a non-reactive polymer material, such as polycarbonate or polypropylene, for example.

Body 1001 defines a plurality of channels, as described with respect to sample cartridge 200 and further below with respect to cartridge 1000. Some of the channels defined in the body 1001 are partially open to facilitate manufacturing by injection molding. Some of the containers defined by body 1001 may be formed with openings to facilitate manufacturing. The cartridge 1000 may include a plurality of membranes connected to the body 1001 to cover and seal some of the container openings and to cover and cooperate to define the channels.

Membranes may be made of any suitable material, such as polypropylene, and may have a thickness ranging, for example, from 20 μm to 200 μm, from 50 μm to 150 μm, or about 100 μm.

The membranes may be secured to the body 1001 by adhesive bonding and/or welding, such as, for example, heat welding or laser welding.

Cartridge 1000 may include:

a base membrane 1402 to cover the channels defined in the base 202;

a waste top membrane 1440 to cover and seal the top of the waste container 240;

a waste side membrane (1442) covering a portion of the waste pneumatic channel (242);

a primary pneumatic side membrane 1412 covering a portion of the primary pneumatic channel 212;

A primary reagent side membrane 1431 covering a portion of the primary reagent channel 231;

a secondary upper membrane (1420) covering and sealing the top of the secondary reagent vessel (220);

a secondary side membrane (1422) covering a portion of the secondary pneumatic channel (222) and secondary reagent channel (232);

a middle outlet membrane 1480 to cover and seal the top of the middle outlet chamber 284T;

QC top membrane 1461 covering and sealing the tops of QC vessel 261 and QC reference vessels 271; and

QC side membrane 1462 covering a portion of the QC pneumatic channel 262 and the QC reference pneumatic channels 272.

Cartridge 1000 has cartridge pneumatic ports 1201, 1202, 1203, 1204, 1205, 1206, 1207, 1208, 1209, 1210 to drive fluid flows through fluid channels in the base and operate cartridge valves. ) and a pneumatic channel plate 1602, defining a plurality of pneumatic channels connecting the pneumatic ports to corresponding pneumatic ports and pneumatic channels of the base 202. The pneumatic channel plate 1602 also has valve ports defined on the base 202 and a valve holder that cooperates with the base membrane 1402 to define the valves, as further described below with respect to FIGS. 7J and 7N. Recesses can be defined.

Pneumatic channel plates 1602 may be coupled to base 202 with base membrane 1402 sandwiched between them. Pneumatic channel plate 1602 may be adhesively bonded to base membrane 1402 and/or base 202 by adhesive bonding, such as a pressure sensitive adhesive (PSA), such as 3M 300LSE adhesive. The adhesive can be prepared as a PSA layer 1606 and define holes corresponding to specific parts that do not require bonding, such as the valves and pneumatic ports of the base 202, for example.

Cartridge 1000 may further include gas permeable membranes configured to allow passage of air but resist or prevent passage of liquid. As discussed with respect to sample cartridge 200, gas permeable membranes can accidentally enter the pneumatic channels of cartridge 100 while escaping cartridge 1000 during processing and potentially contaminating instrument 100. Can be used to restrict liquids.

Gas permeable membranes can be formed of any suitable material for a given application considering the sample and reagents, such as a hydrophobic gas permeable membrane, for example PTFE or PP. Gas permeable membranes can have a thickness of, for example, 20 μm to 200 μm, 50 μm to 150 μm, or about 110 μm.

Cartridge 1000 may include:

a primary permeable membrane 1415 associated with primary pneumatic channel 212 (and also optionally associated with QC, QC pneumatic channel 262 and QC reference pneumatic channels 272);

a secondary permeable membrane (1425) associated with a secondary pneumatic channel (222);

a waste permeable membrane (1445) associated with a waste pneumatic channel (242); and

A mid-outlet permeable membrane (281) associated with the mid-outlet (280).

7D and 7E show the top and bottom surfaces of base 202, base membrane 1402, PSA layer 1606, and pneumatic channel plate 1602 and illustrate how the components are aligned to connect to each other in a stack of layers. do.

Base 202 defines a plurality of channels recessed into the lower surface of base membrane 202, which is covered with base membrane 1402 to define the channels. Base membrane 1402 and PSA layer 1606 define simple sheets with thick holes corresponding to the various ports, valves and holes in base 202. Pneumatic channel plate 1602 defines a plurality of pneumatic channels, valve recesses and cartridge pneumatic ports 1201, 1202, 1203, 1204, 1205, 1206, 1207, 1208 on the upper surface of pneumatic channel plate 1602. , 1209, 1210) are defined on the lower surface of the pneumatic channel plate 1602.

7F-7K are side views through each of the layers visible underneath the cartridge 1000 to facilitate comparison between the layers to illustrate the paths and connections between the vessels, channels, valves and ports. Cross sections are shown. FIG. 7L shows the base 202 (FIG. 7G) overlaid on the channels, valve recesses and pneumatic ports of the pneumatic channel plate 1602 (FIG. 7J) for direct comparison and to view the alignment of the corresponding features. Channels and valves are shown.

Figure 7F shows the top layer of base 202 just above the base channels. Edges 1220 and resilient clips 1222 of the base 202 are shown, configured to engage the rails 1120 and clip recesses 1122 of the cartridge sockets 120. do.

The base may also define a recessed portion 1230 around the reagent vessel 230 with a reduced thickness to mitigate against warping due to material shrinkage after injection molding. For example, base 202 may have a thickness of approximately 2.5 mm at other areas, reducing to approximately 1.5 mm at recessed portion 1230.

Figure 7g shows channels formed in the lower surface of base 202. Referring to FIGS. 7F and 7G, these channels will be described corresponding to the flow schematic of FIG. 2G for illustrative purposes only. The channels and valves may be arranged differently in different embodiments and may include different channels and valves associated with different combinations of vessels.

Primary waste channel 214 is connected to an outlet at the bottom of primary reaction vessel 210 and waste vessel 240 through primary waste valve 215, shown as a gap between two valve ports in FIG. 7G. It extends between the inlets at the bottom.

The secondary waste channel 224 is connected to an outlet at the bottom of the secondary reaction vessel 220 and the waste vessel 240 through the secondary waste valve 225, shown as the gap between the two valve ports in FIG. 7G. It extends between the inlets at the bottom.

The primary reagent channel 231 is connected to the outlet at the bottom of the reagent vessel 230 and the primary reaction vessel 210 ( 7m) extends between the entrances.

As previously discussed, the inlet of primary reagent channel 231 may open into primary reaction vessel 210 near the top of primary reaction vessel 210, as shown in Figure 7M; This is explained in detail in 7o.

In some embodiments, the primary reaction vessel 210 may define an inlet recess 1231 to reduce splashing of liquid reagents flowing into the primary reaction vessel 210. The primary reagent channel 231 may be open facing the first side wall 1231a of the inlet recess 1231 and the opposing second side wall 1231b of the inlet recess 1231. Inlet recess 1231 may be defined in part by a convex surface 1231c between first and second side walls 1231a and 1231b. Liquid reagents entering the inlet recess 1231 at a relatively high velocity may impact the second side wall 1231b and then flow down the convex surface 1231c and down the sidewall of the primary reaction vessel 210. Convex surface 1231c can smoothly transition between the inlet recess 1231 and the sidewall of primary reaction vessel 210. These features can reduce or mitigate against splashing of reagents in the primary reaction vessel 210, which would otherwise allow residual fluids to remain on the side walls or potentially be drawn into the pneumatic channel 212.

Primary reaction vessel 210 may define a similar recess at the opening to primary pneumatic channel 212 to reduce the likelihood of liquid being drawn into primary pneumatic channel 212.

The secondary reagent channel 232 is connected to the outlet at the bottom of the reagent vessel 230 and the secondary reaction vessel 220 ( 7m) extends between the entrances.

As previously discussed, the inlet of secondary reagent channel 232 opens into secondary reaction vessel 220 near the top of secondary reaction vessel 220, as shown in more detail in Figures 7M and 7Q. can do.

In some embodiments, secondary reaction vessel 230 may define an inlet recess similar to inlet recess 1231 of primary reaction vessel 210. In some embodiments, secondary reagent channel 232 may open directly into secondary reaction vessel 220. In some embodiments, secondary reaction vessel 220 may define a concave surface 1232c leading from secondary reagent channel 232 to secondary reaction vessel 220. Concave surface 1232c can smoothly transition between the secondary reagent channel 232 and the sidewall of secondary reaction vessel 220.

Referring to FIGS. 7O and 7P, the lid 211 of the primary reaction vessel 210 is formed integrally with the main body 1001 and is connected to the primary reaction vessel (211a) through a flexible hinge portion 211a. 210). Lid 211 may have a press-fit, snap-fit or interference fit connection, for example, to seal primary reaction vessel 210. It may include an elastically deformable annular flange 1211 configured to engage the upper rim of the primary reaction vessel 210.

References herein to sealed vessels, chambers or channels will be understood to mean sealed other than the defined inlets and outlets for the various ports and channels. In some cases, a fluid tight or air tight seal may be formed. However, depending on the application, a perfect seal may not be necessary and there may be some fluid leakage.

Secondary reagent channel 232 and secondary pneumatic channel 222 may be partially defined in upper surface 1223 surrounding the top of secondary reaction vessel 230, as shown in Figure 7q, and can be relatively It can be opened to a nearby secondary reaction vessel 230.

In some embodiments, the opening of secondary reagent channel 232 to secondary reaction vessel 230 may be relatively distant from the opening of secondary pneumatic channel 222 to secondary reaction vessel 230. For example, the openings may be separated by a distance ranging from 2 mm to 20 mm, 3 mm to 10 mm, 5 mm to 8 mm, or at least 2 mm, at least 3 mm, at least 5 mm, or at least 10 mm.

Secondary reagent channel 232 may extend partially around the circumference of secondary reaction vessel 230 at upper surface 1223 to achieve separation between the openings, for example, as shown in Figure 7R. You can. Alternatively, or additionally, primary pneumatic channel 222 may extend partially around the circumference of secondary reaction vessel 230 at upper surface 1223.

This arrangement may mitigate or reduce accidental aspiration of reagents entering the secondary reaction vessel 220 from the secondary reagent channel 232 into the secondary pneumatic channel 222.

Primary output channel 216 exits primary reaction vessel 210 to join secondary reagent channel 232 through primary output valve 217, shown as the gap between the two valve ports in FIG. 7G. It extends from the outlet at the bottom.

The secondary output channel 226 exits the buffer junction at the bottom of the secondary reaction vessel 220 through the secondary output valve 227, shown as the gap between the two valve ports in Figure 7g. It is extended to (228).

From buffer junction 228, volumetric metering and QC module 260 may include similar channels and valves as described in conjunction with the close-up in Figure 2F, but these are as shown in Figure 7G. , can be arranged differently.

In particular, the base 202 has reference buffer channels ( 1275) is defined. This allows the QC pneumatic channel 262 and the QC reference pneumatic channels 272 to be connected to a single pneumatic line 515 while allowing the QC reference pneumatic channels 272 to be blocked by closing the reference damping valves 1277. do.

The final output channel 256 is connected to a fluid transfer device 880, shown in FIGS. 8A-8C, which provides fluid communication between the final output channel 256 and the output vessel pneumatic port 253 and the output vessel 250. It terminates at a fluid connection port 1256 adjacent to the output vessel pneumatic port 253 and a mechanical clip 1280 configured to connect.

A portion of the waste pneumatic channel 242 extends up the side of the waste container 240, as shown in Figure 7S, and extends down to connect to the waste fluid trap 1640, as shown in Figure 7J. It is shown. Waste fluid trap 1640 includes a chamber defined by base membrane 1402 and a recess 1642 in pneumatic channel plate 1602.

Recess 1642 defines an annular ledge 1643 set at a level above the lower surface 1644 of recess 1642. Waste pneumatic channel 242 is connected to a pneumatic channel plate for fluid communication with recess 1642 connecting through the side wall of recess 1642 above the level of ledge 1643, as shown in FIGS. 7J and 7T. It extends through part of (1602). Bottom surface 1644 defines a hole forming cartridge pneumatic port 1205.

Waste permeable membrane 1445 is secured to ledge 1643 (e.g., by adhesive or heat welding) to isolate cartridge pneumatic port 1205 from waste pneumatic channel 242. Any fluid accidentally drawn in is captured in the waste fluid trap 1640 and restricted from passing through the waste permeable membrane 1445 to the cartridge pneumatic port 1205.

Waste fluid trap 1640 may further include spacer protrusions 1645 extending from lower surface 1644 to the level of ledge 1643 to support waste permeable membrane 1445.

Portions of the primary and secondary pneumatic channels 212, 222 extend over the sides of the primary and secondary reaction vessels 210, 220 in Figure 7F and down toward the pneumatic channel plate 1602 in Figure 7G. It is shown to be. And the QC pneumatic channel 262 and the QC reference pneumatic channels 272 are connected to the pneumatic channel plate 1602 through the channels shown in FIG. 7G, over the sides of the QC vessel 261 and QC reference vessels 271 in FIG. 7F. ) is shown extending downward toward.

Similar to waste fluid trap 1640, primary and secondary pneumatic channels 212, 222 connect to primary and secondary fluid traps 1611, 1620, respectively. QC pneumatic channel 262 and QC reference pneumatic channels 272 also connect to primary fluid trap 1611, as shown in FIG. 7J.

Primary fluid trap 1611 includes a chamber defined by base membrane 1402 and a recess 1612 in pneumatic channel plate 1602.

Recess 1612 defines an annular ledge 1613 set at a level above the lower surface 1614 of recess 1612. Primary pneumatic channel 210 is in fluid communication with recess 1612 connecting through the side wall of recess 1612 above the level of ledge 1613, as shown in FIGS. 7J and 7T. It extends through a portion of plate 1602. Bottom surface 1614 defines a hole forming cartridge pneumatic port 1206.

Primary permeable membrane 1415 is secured to ledge 1613 (e.g., by adhesive or heat welding) to isolate cartridge pneumatic port 1206 from primary pneumatic channel 210, Any fluid accidentally aspirated at 210 is captured in the primary fluid trap 1611 and restricted from passing through the primary permeable membrane 1415 to the cartridge pneumatic port 1206.

Similarly, as shown in FIGS. 7J and 7T, QC pneumatic channel 262 and QC reference pneumatic channels 272 both connect to a common QC pneumatic channel 1660 in pneumatic channel plate 1602, which It connects through the side wall of recess 1612 above the level of 1613. Any fluid accidentally aspirated in the QC pneumatic channel 262 or QC reference pneumatic channels 272 is captured in the primary fluid trap 1611 and flows through the primary permeable membrane 1415 to the cartridge pneumatic port 1206. restricted from passing through.

The primary fluid trap 1611 may further include spacer projections 1615 extending from the lower surface 1614 to the level of the ledge 1613 to support the primary permeable membrane 1415.

Similar to waste and primary fluid traps 1640, 1611, secondary fluid trap 1620 includes a chamber defined by base membrane 1402 and a recess 1622 in pneumatic channel plate 1602. .

Recess 1622 defines an annular ledge 1623 set at a level above the lower surface 1624 of recess 1622. Secondary pneumatic channel 220 is in fluid communication with recess 1622 connecting through the side wall of recess 1622 above the level of ledge 1623, as shown in FIGS. 7J and 7U. It extends through a portion of plate 1602. Bottom surface 1624 defines a hole forming cartridge pneumatic port 1208.

Secondary permeable membrane 1425 is secured to ledge 1623 (e.g., by adhesive or heat welding) to isolate cartridge pneumatic port 1208 from secondary pneumatic channel 220, Any fluid accidentally aspirated at 220 is captured in the secondary fluid trap 1620 and restricted from passing through the secondary permeable membrane 1425 to the cartridge pneumatic port 1208.

The secondary fluid trap 1620 may further include spacer projections 1625 extending from the lower surface 1624 to the level of the ledge 1623 to support the secondary permeable membrane 1425.

The shape, depth, diameter and volume of the fluid traps 1611, 1620, 1640 (as well as the exposed area of the gas permeable membranes 1415, 1425, 1445) determine the amount of liquid that may be accidentally drawn from the pneumatic channels. It can be adjusted to suit different applications. The volume of the fluid traps on the membranes can range, for example, from 10 μL to 1 mL, 50 μL to 500 μL, or about 100 μL, and the surface area of the membranes can range, for example, from 50 mm² to 500 mm², 100 mm² to 300 mm², or about 200 mm². It can be.

Alternatively, in some embodiments, other types of fluid traps may be used, such as microfluidic or gravity type fluid traps. In some embodiments, fluid traps may be omitted entirely.

Referring to Figure 7S, a portion of the waste pneumatic channel 242 may be defined on the exterior side of the waste container 240 and sealed by the waste side membrane 1442. The top of the waste container 240 may be sealed by a waste top membrane 1440.

Each portion of the primary pneumatic channel 212, primary reagent channel 231, secondary pneumatic channel 222, and secondary reagent channel 232 is a support web, as shown in FIG. 7S. 203) (which extends between waste vessel 240, primary reaction vessel 210, reagent vessel 230 and secondary reaction vessel 220) defined on the outer side surface and corresponding side membranes 1412 , 1431, 1422). Other portions of the channels 212, 231, 222, 232, for example, near the base 202 and/or near the tops of the reaction vessels 210, 220, as shown in FIGS. 7M and 7Q. It can be completely defined by the body 1001.

A portion of secondary pneumatic channel 222 and secondary reagent channel 232 may be defined in upper surface 1223 and sealed by secondary upper membrane 1420, as shown in FIGS. 7Q and 7R.

The sealing membranes may include separate parts 1401 to facilitate assembly, which can be removed once the corresponding sealing membrane is secured to the body 1001.

Referring to Figure 7v, the QC pneumatic channel 262 and a portion of the QC reference pneumatic channels 272 are defined on the outer side of the QC vessel 261 and the QC reference vessels 271, and the QC side membrane 1462 It can be sealed by. And the QC top membrane 1461 may be provided to cover and seal the tops of the QC container 261 and the QC reference containers 271.

The middle outlet membrane 1480 may be secured to the upper surface of the base 202 to cover and seal the top of the middle outlet chamber 284. This is shown in more detail in Figures 7w and 7x.

Intermediate outlet chamber 284 may be defined by a recess in the upper surface of base 202. The lower surface of outlet chamber 284 may define openings to intermediate outlet 280 and intermediate outlet pneumatic channel 282. The mid-outlet permeable membrane 281 may be secured (glued or heat-welded) to the lower surface of the mid-outlet chamber 284 to cover the mid-outlet 280, but not the mid-outlet pneumatic channel 282. The upper opening of middle outlet chamber 284 may be sealed by middle outlet membrane 1480.

When the pneumatic module 500 is operated to reduce the pressure in the intermediate outlet pneumatic channel 282, air is forced into the measurement channel such that the liquid in the measurement channel is drawn up to the membrane 281 without passing into the chamber 284. It may be drawn through the permeable membrane 281 from 299 (and/or outlet channel 285).

Referring to Figure 7J, pneumatic channel plate 1602 also defines a plurality of pneumatic channels connecting other pneumatic ports to the valve recesses to allow operation of the cartridge valves. Cartridge valves include switch valves configured as shown in FIG. 2K.

For example, a vertical cross-section of primary waste valve 215 is shown in Figure 7N. A primary waste channel 214 may be defined in the base 202 extending from an outlet at the bottom of the primary reaction vessel 210 towards the waste vessel 240 . There may be a break in the primary waste channel 214 which is separated at valve 215 into two sections terminating at first and second valve ports 214a and 214b.

Valve ports 214a, 214b may be closed in a rest configuration by base membrane 1402 (which also closes the corresponding valve ports of all other cartridge valves in a rest configuration). On the other side of base membrane 1402, opposite valve ports 214a, 214b, is defined by pneumatic channel plate 1602 and valve ports 214a, 214b (opposite side of base membrane 1402). There is a primary waste valve recess 1615 extending therebetween.

Primary waste valve recess 1615 can be connected to cartridge pneumatic port 1204 such that pneumatic module 500 can be operated to reduce the pressure in primary waste valve recess 1615, Deflection of the base membrane 1402 toward the valve recess 1615 causes the primary waste valve 215 to open and allow fluid communication between the first and second valve ports 214a, 214b. In some embodiments, positive pressure is applied to the valve recess 1615 (and others) such that it remains closed, for example, during operations that may apply pressure to the valve to open it. It can be.

In a similar manner, other cartridge valves may include valve ports and corresponding valve recesses connected to cartridge pneumatic ports by pneumatic channels to allow operation of the valves by pneumatic module 500.

Pneumatic channel plate 1602 defines a primary waste valve recess 1615 that corresponds to primary waste valve 215 and is connected to cartridge pneumatic port 1204 via pneumatic channel 1604.

Pneumatic channel plate 1602 defines a secondary reagent valve recess 1636 that corresponds to secondary reagent valve 236 and is connected to cartridge pneumatic port 1204 via pneumatic channel 1604.

Pneumatic channel plate 1602 defines a primary output valve recess 1617 that corresponds to primary output valve 217 and is connected to cartridge pneumatic port 1207 via pneumatic channel 1607.

Pneumatic channel plate 1602 corresponds to primary reagent valve 235 and defines primary reagent valve recess 1635 connected to cartridge pneumatic port 1203 via pneumatic channel 1603.

Pneumatic channel plate 1602 defines a secondary waste valve recess 1625 that corresponds to secondary waste valve 225 and is connected to cartridge pneumatic port 1203 via pneumatic channel 1603.

Pneumatic channel plate 1602 defines a secondary outlet valve recess 1627 that corresponds to secondary outlet valve 227 and is connected to cartridge pneumatic port 1209 via pneumatic channel 1609.

Pneumatic channel plate 1602 defines a final output valve recess 1657 that corresponds to final output valve 257 and is connected to cartridge pneumatic port 1209 via pneumatic channel 1609.

Pneumatic channel plate 1602 defines three reference buffer valve recesses 1677 that correspond to three reference buffer valves 1277 and are connected to cartridge pneumatic port 1209 via pneumatic channel 1609.

Pneumatic channel plate 1602 defines a QC vessel valve recess 1664 that corresponds to QC vessel valve 264 and is connected to cartridge pneumatic port 1210 via pneumatic channel 1610.

Pneumatic channel plate 1602 defines a QC buffer valve recess 1667 that corresponds to QC buffer valve 267 and is connected to cartridge pneumatic port 1210 via pneumatic channel 1610.

Valve recesses may include any suitable size, shape and proportion for a particular application. For example, valve recesses may be generally rectangular with rounded corners, as shown in the figures. The valve recesses of the illustrated embodiment are all similar in dimensions, for example having a length of about 6 mm, a width of about 2 mm and a depth of about 0.5 mm.

The pneumatic channels defined in pneumatic channel plate 1602 are approximately 0.5 mm deep and approximately 1 mm wide.

Similarly, the fluid channels may be about 0.5 mm deep and about 1 mm wide. However, some of the fluid channels may have smaller dimensions, for example the volumetric section, where the smaller channel cross-section allows more precise control of the volumetric flow rate and the volume of liquid in the channels.

For example, channels 226, 256, 266, 269, 299, 285 may have a width of less than 0.5 mm, less than 0.3 mm, or about 0.25 mm and a depth of less than 1 mm, less than 0.5 mm, less than 0.3 mm, or about 0.2 mm. You can have These channels may be flared to form valve ports for each valve with a width of approximately 1 mm, for example, as shown in FIG. 7x.

In some embodiments, all valve ports are as shown in Figure 7Y, which illustrates a welding pattern for welding base membrane 1402 to base 202 (e.g., by heat welding or laser welding). , may arise from corresponding channels. Some suitable dimensions for the different valves of cartridge 1000 are provided in the enlarged views of Figure 7Y for illustrative purposes only. The corresponding valve recesses of the pneumatic channel plate 1602 can substantially match the shape and dimensions shown in FIG. 7Y, for example.

The weld pattern may include internal weld lines 1403 that run along the edges of the channels and connect valve ports to form valves. In some embodiments, the weld pattern may also include external weld lines 1404 that provide a second barrier around the channels for redundancy.

Referring to FIG. 7Z, measurement channel 299 is shown in more detail with close-ups of buffer joint 228 and quality control joint 229, according to some embodiments. Channel and joint shapes can include any suitable geometry for a given application. For example, in Figure F the joints 228 and 229 are shown as vertical T-junctions. Alternatively, as shown in FIG. 7Z, joints 228, 229 may include, for example, angled Y-junctions, or curved Y-junctions, or any other suitable geometry.

를 형성할 수 있다.At quality control junction 229, quality control channel 269 is at an obtuse angle with measurement channel 299.

can be formed.

를 형성할 수 있다.At the buffer junction 228, the buffer channel 266 is at an obtuse angle with the measurement channel 299.

can be formed.

을 형성할 수 있다.Similarly, at quality control junction 229, output channel 285 is at an obtuse angle with measurement channel 299.

can be formed.

을 형성할 수 있다.At buffer junction 228, secondary output channel 226 (or alternatively, primary output channel 216) is at an obtuse angle with measurement channel 299.

can be formed.

,

,

,

의 각각은 예를 들어, 90° 내지 180°, 100° 내지 170°, 110° 내지 160°, 120° 내지 150°, 130° 내지 140°, 약 135° 또는 약 137°의 범위일 수 있다.joint angles

,

,

,

Each of can range, for example, from 90° to 180°, 100° to 170°, 110° to 160°, 120° to 150°, 130° to 140°, about 135°, or about 137°.

Measurement channels 299 may splay at each junction 228, 229 and define curved edges that transition into connecting channels 226, 266, 269, 285.

과 변곡점(229a)을 형성할 수 있다.At quality control junction 229, quality control channel 269 and output channel 285 are at an acute angle.

and an inflection point 229a may be formed.

및 변곡점(228a)을 형성할 수 있다.At buffer junction 228, buffer channel 266 and secondary output channel 226 (or alternatively, primary output channel 216) form an acute angle.

and an inflection point 228a may be formed.

및

의 각각은 예를 들어, 10° 내지 90°, 30° 내지 60°, 약 45° 또는 약 40°의 범위일 수 있다.acute angle

and

Each of can range, for example, from 10° to 90°, from 30° to 60°, from about 45°, or from about 40°.

The radius of curvature of the inflection points 228a and 229a may range, for example, from 0.1 mm to 0.5 mm, less than 0.5 mm, less than 0.4 mm, less than 0.3 mm, less than 0.2 mm, or about 0.2 mm.

Referring to FIG. 7H, the base membrane 1402 is between the base 202 and corresponding portions of the ports and channels 242, 212, 222, 253, 283, 262, and 272 of the pneumatic channel plate 1602. Holes are defined to allow fluid communication through the base membrane 1402. Base membrane 1402 also includes holes 1230 that allow passage of the radiator and magnets.

In the QC and measurement portion of cartridge 1000, intermediate outlet 280 is connected to the cartridge pneumatic port (282) through intermediate outlet pneumatic channel 282, defined in pneumatic channel plate 1602, as shown in FIGS. 7J and 7X. 1202).

Output vessel pneumatic port 253 is connected to cartridge pneumatic port 1201 via output vessel pneumatic channel 252, which may be defined in pneumatic channel plate 1602, as shown in FIG. 7J.

Pneumatic channel plate 1602 defines a QC hole 1261 aligned with QC vessel 261 and three QC reference holes 1271 aligned with QC reference vessels 271. Polypropylene membrane 1402 is used to analyze the contents of QC vessel 261 and reference vessels 271 through QC aperture 1261 and QC reference apertures 1271. 271) and may provide a transparent viewing window allowing optical access to the optical module.

In some embodiments, pneumatic channel plate 1602 (or alternatively another portion of cartridge 1000) may activate a switch or switch on device 100 to indicate that cartridge 1000 is correctly installed in socket 120. A switch recess 1690 configured to engage a microswitch may be defined.

FIG. 7I illustrates the PSA layer 1606 in more detail and shows holes corresponding to areas that do not require adhesive bonding, such as valve recesses, fluid traps, and QC hole 1261. and all recesses in the upper surface of the pneumatic channel plate 1602, including the QC reference holes 1271 and the holes in the base membrane 1402. The PSA layer 1606 does not need to define holes for the pneumatic channels of the pneumatic channel plate since the adhesive may not affect the function of the channels.

8A and 8B, fluid transfer device 880 is shown in greater detail. Device 880 includes a temporary removable lid 259 used to seal output container 250 closed during instrument workflow as described with respect to FIGS. 2F and 2G.

Apparatus 880 includes a delivery pneumatic channel 882 configured to connect to fluidically connect output vessel 250 to output vessel pneumatic channel 252, and a temporary lid 259 to transfer liquid from final output channel 256. It further includes a liquid delivery channel 886 configured to convey to the output container 250 through the outlet of.

Channels 882, 886 are defined in body 888 (e.g., injection molded polypropylene) and transport device membrane 840 (e.g., heat-welded polypropylene membrane) as shown in FIG. 7C. ) and sealed.

Body 888 also defines a connector 889 configured to mechanically couple to a corresponding connector 1289 on the upper surface of base 202 near output container seat 254, thereby as shown in Figure 8C. , connecting the delivery pneumatic channel 882 to the output vessel pneumatic channel 252 and the liquid delivery channel 886 to the final output channel 256.

Body 888 may be resiliently flexible and, when bent into a twisted connection configuration, as shown in Figure 7A, may fluidly and mechanically couple output container 250 to cartridge 1000; Body 888 presses output container 250 toward cartridge 1000 (e.g., into sheet 254) to secure the cartridge during instrument operations, such as mixing with an orbital shaker. It can be configured to do so.

Fluid transfer device 880 may be provided in a kit, for example, with cartridge 1000 (and also optionally output container 250).

Alternatively, temporary lid 259 may simply be connected to channels 256, 252 via tubes rather than to fluid transfer device 880.

Cartridge 1000 may include markings 1295, such as a barcode, to identify samples stored therein (FIG. 7A). Cartridge 1000 may be provided with an output container 250 that may include a corresponding indicia 1296 or barcode that may be the same as or related to indicia 1295. Alternatively, cartridge 1000 may include one or more peel off labels with corresponding indicia 1296, which can be removed from cartridge 1000 and appropriate for receiving the output fluid once the sample has been processed. It can be applied to the output container 250.

Indications 1295, 1296 may be scanned or input data into a laboratory information system, etc. to be associated with data generated by device 1000 when processing a sample.

Example 1

For illustrative purposes only an example of an instrument workflow will now be described. In some embodiments, instrument 100 may be configured to perform, for example, a nucleic acid extraction workflow.

Before the instrument workflow begins, a user may pipet a fluid sample, such as a biological specimen, into the primary reaction vessel 210 of the sample cartridge 200. For example, 0.2 to 5 mL of blood or bone marrow collected from a patient.

The user then closes the lid 211 of the primary reaction vessel 210 and then records or scans the serial number or other markings on the sample cartridge 200, for example from a vial previously containing a sample. Corresponding patient details can be recorded. This information may be recorded, for example, in a LIMS system or laboratory information system.

The user can then insert the cartridge 200 into one of the cartridge slots 120 of the device 100.

The user can then use the user interface to select a workflow program for the device. The device workflow may then begin with device functions controlled by the control module 101 according to instructions recorded on computer-readable storage media. For example, the nucleic acid extraction workflow described below.

The motion module is operative to engage the pneumatic module with the pneumatic ports of the sample cartridge and clamp the cartridge to limit removal of the cartridge 200 from the cartridge slot 120.

The motion module is then operated to move the reagent module into position above the sample cartridge and the reagent module is operated to dispense Proteinase K into the reagent vessel 230. For example, ranging from 50 to 100 μg of proteinase K per mL of sample volume.

The pneumatic module is operated to transfer the reagent along with the sample to the primary reaction vessel 210.

The mixing module's orbital shaker operates to promote mixing of the sample and reagents in the primary reaction vessel.

The motion module and heat module are operated to activate and raise the heater to heat the primary reaction vessel and incubate it at 62 C for 10 minutes to digest the proteins in the blood. The heater can then be lowered and deactivated.

The motion module and the reagent module are then operated to dispense lysis buffer (e.g., 5M Guanadinium HCl, 0.25% Tween-20) into the reagent container 230.

Then, the pneumatic module is operated to transfer the lysis buffer to the primary reaction vessel.

The motion module and reagent module are then operated to dispense functionalized magnetic beads (e.g. Carboxyl COOH Magbeads) into the reagent vessel.

Any suitable type of functionalized bead may be used, including: for example solid phase reversible immobilization (SPRI) functionalized beads, carboxylated beads, or other self-functionalized beads. .

The pneumatic module is then activated to transfer the beads to the primary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the primary reaction vessel.

The motion and heat modules are operated to heat the primary reaction vessel and incubate the contents at 62 C for 15 minutes to lyse the blood and assemble the nucleic acid (NA) into the beads.

Then, the heater is deactivated and the cooling fan is operated to cool the primary reaction vessel 210.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel containing the lysate to the waste vessel 240.

The magnets 710 are then disengaged from the primary reaction vessel.

The motion module and reagent module are then operated to dispense Wash 1 buffer (e.g., 3M guanadinium HCl, 30% ethanol) into the reagent vessel.

The pneumatic module is then actuated to transfer the Wash 1 solution to the primary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense wash 2 buffer (e.g., 20mM glycine.HCl (pH 3.0) 80% ethanol) into the reagent vessel.

The pneumatic module is then actuated to transfer the wash 2 solution to the primary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense Wash 3 buffer (e.g., 20mM Glycine.HCl(pH 3.0)+0.1% Tween 20) into the reagent vessel.

The pneumatic module then operates to transfer the wash 3 solution to the primary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is activated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense Wash 4 buffer (e.g., 20mM Glycine.HCl (pH 3.0) +0.1% Tween 20) into the reagent vessel.

The pneumatic module is then actuated to transfer the wash 4 solution to the primary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and the reagent module are then operated to dispense the elution buffer (e.g., 1xTE, pH8.0) into the reagent vessel.

Then, the pneumatic module is operated to transfer the elution buffer to the primary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the primary reaction vessel.

The heater is raised and activated to heat the primary reaction vessel to 74 C for 15 minutes to release the DNA from the beads into the elution buffer.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents (effluent) of the primary reaction vessel to the secondary reaction vessel 220.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then operated to dispense the COOH (carboxyl) beads into the reagent vessel with combination buffer (e.g., 0.8M NaCl + 11% PEG8000).

The pneumatic module is then activated to transfer the contents of the reagent vessel to the secondary reaction vessel.

The orbital shaker is operated to facilitate mixing of the secondary reaction vessel contents and assembly of the extracted DNA to the COOH beads.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the secondary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the secondary reaction vessel to the waste container 240.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense COOH bead wash 1 (e.g., 85% ethanol) into the reagent vessel.

The pneumatic module is then actuated to transfer the contents of the reagent vessel to the secondary reaction vessel.

The contents of the secondary reaction vessel are then allowed to incubate for 30 seconds.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the secondary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the secondary reaction vessel to the waste container 240.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then operated to dispense COOH bead wash 2 (e.g., 85% ethanol) into the reagent vessel.

The pneumatic module is then actuated to transfer the contents of the reagent vessel to the secondary reaction vessel.

The contents of the secondary reaction vessel are then allowed to incubate for 30 seconds.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the secondary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the secondary reaction vessel to the waste container 240. The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then operated to dispense COOH elution buffer (e.g., 10mM Tris, pH 8.0) into the reagent vessel.

The pneumatic module is then actuated to transfer the elution buffer to the secondary reaction vessel.

The orbital shaker is operated to promote mixing of the contents of the secondary reaction vessel to release DNA from the COOH beads into the elution buffer.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the secondary reaction vessel.

The pneumatic module is then actuated to draw the liquid contents (effluent) of the secondary reaction vessel up to the air-permeable membrane and fill the metering channel.

The motion module and the reagent module are then operated to distribute the neutral buffer to the QC buffer vessel 265 and the three QC reference buffer vessels 275.

The pneumatic module is then actuated to draw the buffer from the QC buffer vessel through the metering channel into the QC vessel along with an aliquot (e.g., 1 μL) of the eluate in the metering channel. Then, the buffer solution is transferred from the QC reference buffer containers 275 to the corresponding QC reference containers 271.

The pneumatic module is then operated to transfer the remaining eluate from the secondary reaction vessel to the output vessel 250.

The orbital shaker is operated to facilitate mixing of the contents of the QC vessel 261 and the QC reference vessel 271 and to facilitate resuspension of the dye and reference nucleic acid (NA) preloaded in the QC reference vessel.

The motion module is operative to move the optical module to a position corresponding to a QC vessel containing an aliquot of eluate along with the sample cartridge and buffer, and the optical module is operative to perform fluorescence measurements on the contents of the QC vessel.

The motion module is further operated to move the optical module to three positions corresponding to the three QC reference containers, and the optical module is operated to perform fluorescence measurements on the contents of each of the QC reference containers.

Fluorometric data is then used to determine the DNA concentration of the final eluate. The resulting data provides quantification and can be transmitted to a LIMS system for recording and/or capturing.

Finally, the pneumatic module can be lowered and separated from the sample cartridge. This may include the end of a workflow program according to some embodiments.

Sample cartridge 200 can then be removed from device 100 by the user. The temporary lid 259 can be removed from the output container 250 and the main lid can be closed to seal the output container 250.

Output container 250 can then be removed from output container sheet 254 and the remainder of sample cartridge 200 can be discarded.

Example 2

Another workflow example according to some embodiments is described below. Chemical and operating parameters are appropriate for gDNA extraction from 0.5 mL whole blood samples. Details of device operation may also be relevant to other applications and processes.

Before the instrument workflow begins, a user may pipet a fluid sample, such as a biological specimen, into the primary reaction vessel 210 of the sample cartridge 200. For example, 0.5 mL of blood taken from a patient.

The user then closes the lid 211 of the primary reaction vessel 210 and then records or scans the serial number or other markings on the sample cartridge 200 and retrieves corresponding patient details, for example from a vial previously containing a sample. Information can be recorded. This information may be recorded, for example, in a LIMS system or laboratory information system.

The user can then insert the cartridge 200 into one of the cartridge slots 120 of the device 100.

The user can then use the user interface to select a workflow program for the device. The device workflow may then begin with device functions controlled by the control module 101 according to instructions recorded on computer-readable storage media. For example, the nucleic acid extraction workflow is described below.

The motion module is operated to engage the pneumatic module with the pneumatic ports of the sample cartridge and clamp the cartridge to limit removal of the cartridge 200 from the cartridge slot 120.

The motion module is then actuated to move the reagent module into position above the sample cartridge and the reagent module is actuated to dispense 50 μL of Proteinase K (Qiagen, as received from the supplier) into the reagent vessel 230. .

The pneumatic module is operated to transfer the reagent along with the sample to the primary reaction vessel 210.

The reagent module is then actuated to dispense 120 μL of commercial support buffer AL into the reagent vessel (230) and the pneumatic module is actuated to transfer the reagent along with the specimen to the primary reaction vessel (210).

Alternatively, proteinase K and buffer may be dispensed together or sequentially in reagent vessel 230 and then transferred together to primary reaction vessel 210 in a single transfer step.

Operation of the pneumatic module may include applying vacuum pressure or negative pressure (relative to ambient pressure), for example in the range of 100 mBar to 120 mBar.

The orbital shaker in the mixing module operates at 1100 rpm for 10 seconds to promote mixing of samples and reagents in the primary reaction vessel.

The motion module and heat module are operated to activate and raise the heater to heat the primary reaction vessel and incubate it at 25°C for 10 minutes to digest the proteins in the blood. The heater can then be lowered and deactivated.

Alternatively, if the ambient temperature is close to 25°C, heaters may not be required for this step.

The motion module and reagent module then dispense 825 μL elution buffer (0.8M g.HCl, 0.01M Tris pH8, 50% 2-propanol, 1.2M NaCl, 2mM EDTA, 0.25% Tween-20) into the reagent vessel 230. It operates so that

The pneumatic module is then actuated to transfer the elution buffer to the primary reaction vessel.

The motion module and reagent module are then operated to dispense functionalized magnetic beads (Siemens Versant 50 μL) into the reagent vessel.

The pneumatic module is then actuated to transfer the beads to the primary reaction vessel.

To avoid or reduce the time available for the beads to precipitate or sediment in the reagent vessel (which can cause clogging), the pneumatic module places the beads in a primary chamber before completing their dispensing into the reagent vessel. It can be operated to transfer to a reaction vessel. For example, transfers may begin during or during distribution and may be performed in stages. Dispensing may also be performed in stages in some embodiments.

Alternatively or additionally, a portion (e.g., 2/3) of the Lysis buffer is retained and transferred to the reagent chamber after dispensing and transferring the beads to wash away beads remaining in the reagent vessel or to transfer the channel to the primary reaction vessel. can be distributed.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the primary reaction vessel.

The motion and heat modules are operated to heat the primary reaction vessel and incubate the contents at approximately 62°C for 15 minutes to lyse the blood and assemble the nucleic acids (NA) into the beads. The orbital shaker can also be operated at 1100 rpm during the incubation period to promote mixing.

The heater is then deactivated and the cooling fan is turned on to cool the primary reaction vessel 210 back to ambient temperature. For example, the cooling operation may have a duration ranging from 1 minute to 5 minutes, 2 minutes to 3 minutes, or about 2 minutes depending on the cooling rate. In some embodiments, it may be necessary to cool the reaction vessel to prevent the beads from drying out. In other embodiments, this step may be omitted if drying is not a problem.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The magnets may engage during cooling operation.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel containing the melt to the waste vessel 240. The magnets are applied to engage the beads for about one minute prior to transferring the liquid so that the beads migrate towards the magnets and are held against the wall of the vessel with sufficient strength to prevent them from flowing with the liquid during the transfer process. You can. The length of time required may vary depending on the strength of magnetic attraction between the beads and magnets and the viscosity of the fluid. In some embodiments, a shorter settling time (less than 1 minute) may be sufficient or a longer settling time (e.g., more than 1 minute, more than 2 minutes, more than 3 minutes, or more than 4 minutes) may be necessary.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense 850 μL of Wash 1 buffer (e.g., 3M guanadinium HCl (gHCl), 30% ethanol) into the reagent vessel.

The pneumatic module is then actuated to transfer the Wash 1 solution to the primary reaction vessel.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is activated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240. The magnets can be applied to engage the beads for about 1 minute before transferring the liquid.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense 450 μL of Wash 2 buffer (e.g., 80% ethanol, 0.1 M sodium citrate buffer, pH 3) into the reagent vessel.

The pneumatic module is then actuated to transfer the wash 2 solution to the primary reaction vessel.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240. The magnets can be applied to engage the beads for about 1 minute before transferring the liquid.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense 450 μL of Wash 3 buffer (e.g., 20mM Glycine.HCl, 0.1% Tw-20, pH3) into the reagent vessel.

The pneumatic module is then actuated to transfer the wash 3 solution to the primary reaction vessel.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnet 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240. The magnets can be applied to engage the beads for about 1 minute before transferring the liquid.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then actuated to dispense 450 μL of Wash 4 buffer (e.g., 20mM Glycine.HCl, 0.1% Tw-20, pH3) into the reagent vessel.

Wash 4 is completed using the same buffer as Wash 3 to wash away contaminants from the distribution system from the previous steps. This step may be repeated more than once if necessary to ensure purity or further reduce the likelihood of contaminants appearing in the solution. Alternatively, this step can be omitted if contaminants are not a problem or if the distribution system includes independent channels to avoid potential contamination.

The pneumatic module is then actuated to transfer the wash 4 solution to the primary reaction vessel.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240. The magnets can be applied to engage the beads for about 1 minute before transferring the liquid.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module are then operated to dispense 165 μL of elution buffer into the reagent vessel.

The pneumatic module is then actuated to transfer the elution buffer to the primary reaction vessel.

The heater is raised and activated to heat the primary reaction vessel to approximately 62°C for 10 minutes to release the DNA from the beads into the elution buffer. The orbital shaker may be operated at 1100 rpm for a 10 minute incubation period to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are then operated to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents (effluent) of the primary reaction vessel to the secondary reaction vessel 220. The magnets can be applied to engage the beads for about 1 minute before transferring the liquid. The spent beads then remain in the primary reaction vessel until the end of the process (during further processing of the eluate in the secondary reaction vessel) or until the cartridge is discarded.

The magnets 710 are then separated from the primary reaction vessel.

The motion module and reagent module then combine COOH (carboxyl) beads in a combination buffer (e.g. 470 μL mastermix, 1.24 M NaCl, 13.95% PEG8000, 0.78% w/v magnify (MFY0002 Bangslab beads). s)) is also operated to dispense into reagent containers.

The pneumatic module is then actuated to transfer the contents of the reagent vessel to the secondary reaction vessel.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the primary reaction vessel.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the primary reaction vessel.

The beads are held in place for 1 to 5 minutes while the pneumatic module is operated to transfer the liquid contents of the primary reaction vessel to the waste vessel 240. The magnets can be applied to engage the beads for about 1 minute before transferring the liquid.

In this example, the beads in the secondary reaction vessel beads have a weaker magnetic attraction to the magnets and are in a more viscous solution. Therefore, longer settling times (e.g., 2 minutes) may be required. However, longer or shorter settling times may be used, from less than 1 minute to more than 2, 3 or 4 minutes, if sufficient.

In some embodiments the magnets may remain engaged during subsequent cleaning steps. For example, if the beads have relatively weak assembly kinetics, as in this case, holding the beads in place with magnets can mitigate premature washing of DNA from the beads.

The motion module and reagent module are then actuated to dispense 200 μL of COOH Bead Wash 1 (e.g., 85% ethanol) into the reagent vessel.

The pneumatic module is then actuated to transfer the contents of the reagent vessel to the secondary reaction vessel.

The contents of the secondary reaction vessel are then allowed to incubate for 30 seconds at room temperature.

The pneumatic module is then actuated to transfer the liquid contents of the secondary reaction vessel to the waste vessel 240, while magnets (still engaging) hold the beads in place.

The motion module and reagent module are then operated to dispense 200 μL of COOH Bead Wash 2 (e.g., 85% ethanol) into the reagent vessel.

COOH Bead Wash 2 is completed using the same buffer as COOH Bead Wash 1 to wash away contaminants from the distribution system from the previous steps. This step may be repeated more than once if necessary to ensure purity or further reduce the likelihood of contaminants appearing in the solution. Alternatively, this step can be omitted if contaminants are not a problem or if the distribution system includes independent channels to avoid potential contamination.

The pneumatic module is then actuated to transfer the contents of the reagent vessel to the secondary reaction vessel.

The contents of the secondary reaction vessel are then allowed to incubate for 30 seconds at room temperature.

The pneumatic module is then actuated to transfer the liquid contents of the secondary reaction vessel to the waste vessel 240, while magnets (still engaging) hold the beads in place.

The magnets 710 are then separated from the primary reaction vessel.

The operation module and reagent module are then actuated to dispense 30 μL of COOH elution buffer (e.g. 1x TE buffer pH 8) into the reagent vessel.

The pneumatic module is then actuated to transfer the elution buffer to the secondary reaction vessel.

The orbital shaker was operated at 1100 rpm for 10 seconds to promote mixing of the contents of the secondary reaction vessel and release the DNA from the COOH beads into the elution buffer.

The motion module and magnetic module are operative to engage magnets 710 and retain the magnetic beads to one or more sides of the secondary reaction vessel. A settling time of approximately 1 minute may be allowed before the next step.

The pneumatic module is then actuated to draw the liquid contents (effluent) of the secondary reaction vessel up to the air-permeable membrane and fill the metering channel.

The motion module and the reagent module then add a neutral buffer (e.g., 199 μL 1x TE buffer pH 8) to the QC buffer vessel 265 and three QC reference buffer vessels 275 (e.g., 200 μL 1x TE buffer pH 8). It operates to distribute.

The pneumatic module is then operated to pull buffer from the QC buffer vessel through the metering channel into the QC vessel 265 along with an aliquot (e.g., 1 μL) of eluate from the metering channel until air fills the channels. The pneumatic module is also operated to transfer buffer fluid from the QC reference buffer vessels 275 to the corresponding QC reference vessel 271.

Each of the QC vessel 265 and QC reference buffer vessels 275 contains a similar amount (e.g., 0.2 μg) of dried DNA dye and the QC reference buffer vessels 275 each contain a different reference amount for comparison. of gDNA (e.g., 4 ng gDNA, 60 ng gDNA, and 500 ng gDNA respectively).

The pneumatic module is then operated to transfer the remaining eluate from the secondary reaction vessel to the output vessel 250.

The orbital shaker is operated at 1100 rpm for 10 seconds to facilitate mixing of the contents of the QC vessel 261 and the QC reference vessels 271 and to facilitate resuspension of the pre-loaded dye and reference nucleic acid (NA) in the QC reference vessels. do.

The motion module is operated to move the optical module to a position corresponding to a QC container containing a quantity of eluate along with the sample cartridge and buffer, and the optical module is operated to perform fluorescence measurements on the contents of the QC container.

The motion module is further operated to move the optical module to three positions corresponding to the three QC reference containers, and the optical module is operated to perform fluorescence measurements on the contents of each QC reference container.

Data from the fluorescence measurements are used to determine the DNA concentration of the final eluate by fitting and interpolating a curve between measurements from three reference vessels with known concentrations (or extrapolating to determine the concentration of the eluate). . Resulting data may be transmitted to a LIMS system for recording and/or capture.

Finally, the pneumatic module can be lowered and separated from the sample cartridge. This may include the end of the workflow program according to some embodiments.

Sample cartridge 200 may then be removed from device 100 by the user. The temporary lid 259 can be removed from the output container 250 and the main lid can be closed to seal the output container 250.

Output container 250 can then be removed from output container sheet 254 and the remainder of sample cartridge 200 can be discarded.

Those skilled in the art will understand that various changes and/or modifications may be made to the above-described embodiments without departing from the broad spirit of the present disclosure. Accordingly, the present embodiments should be regarded in all respects as illustrative and not restrictive.

Claims (71)

In a sample cartridge for a chemical processing instrument,
a primary reaction vessel configured to receive a fluid sample for processing and configured to receive a lid for closing an open top of the primary reaction vessel;
a reagent vessel configured to receive one or more fluid reagents through an open top of the reagent vessel, wherein the reagent vessel controls fluid flow through a primary reagent channel; connected to the primary reaction vessel through the primary reagent channel with a primary reagent valve disposed in the channel; and
a pneumatic module in fluid communication with the primary reaction vessel and for selectively adjusting the pressure within the primary reaction vessel when the lid is closed to draw the fluid contents of the reagent vessel into the primary reaction vessel; ), comprising a primary pneumatic port configured to be connected to a sample cartridge. 2. The method of claim 1, further comprising a primary pneumatic channel extending between the primary pneumatic port and the primary reaction vessel,
The sample cartridge of claim 1, wherein the opening of the primary pneumatic channel into the primary reaction vessel is located somewhat above a side wall of the primary reaction vessel. The sample cartridge of claim 2, wherein the opening of the primary pneumatic port to the primary reaction vessel is located closer to the top of the primary reaction vessel than the bottom of the primary reaction vessel. 4. The sample cartridge of any one of claims 1 to 3, wherein the opening of the primary reagent channel to the primary reaction vessel is located somewhat above a side wall of the primary reaction vessel. 4. The sample cartridge of claim 3, wherein the opening of the primary reagent channel to the primary reaction vessel is located closer to the top of the primary reaction vessel than to the bottom of the primary reaction vessel. The sample cartridge of any preceding claim, further comprising a final output channel configured to convey final output fluid from the primary reaction vessel to a removable output vessel. 7. The method of claim 6, configured to be in fluid communication with the output vessel through an output vessel pneumatic channel and to draw the final output fluid from the primary reaction vessel to the output vessel through the final output channel. and an output vessel pneumatic port configured to be connected to a pneumatic module to selectively adjust the pressure in the output vessel. 8. The sample cartridge of claim 7, further comprising a temporary lid configured to close the output container during processing, the temporary lid configured to fluidly connect the final output channel and the output container pneumatic channel to the output container. According to any one of claims 6 to 8,
a sealed quality control vessel configured to receive an aliquot of the output fluid for quality control analysis;
a quality control channel extending between the quality control vessel and a quality control junction with the final output channel; and
a pneumatic module in fluid communication with the quality control vessel and for selectively adjusting the pressure within the quality control vessel to aspirate the aliquot of final output fluid from the final output channel to the quality control vessel through the quality control channel. A sample cartridge further comprising a quality control pneumatic port configured to be connected. 10. The sample cartridge of claim 9, wherein the quality control vessel is pre-loaded with a dye to be mixed with the aliquot of final output fluid for quality control analysis. According to claim 9 or 10,
a buffer solution vessel configured to receive a buffer solution through an open top of the buffer solution vessel for mixing with the final output fluid for quality control analysis;
a buffer channel extending between the buffer channel and the buffer junction with the final output channel between the quality control junction and the primary reaction vessel; and
A sample cartridge further comprising a buffer channel valve disposed in the buffer channel to control the flow of the buffer through the buffer channel. According to any one of claims 9 to 11,
an intermediate outlet from the final output channel between the quality control joint and the output container;
a sealed chamber into which the intermediate outlets are opened;
an air-permeable liquid barrier membrane covering the outlet; and
an intermediate outlet pneumatic port in fluid communication with the sealed chamber and configured to be connected to a pneumatic module to selectively adjust pressure within the sealed chamber to draw air from the final output channel through the air permeable membrane. A sample cartridge further comprising a port). 13. The method of any one of claims 1 to 12, further comprising: a sealed waste container configured to receive waste fluid from the primary reaction vessel through a waste channel; and
A waste pneumatic port configured to be in fluid communication with the waste container and connected to a pneumatic module to selectively adjust pressure within the waste container to draw fluid from the primary reaction vessel to the waste container through the waste channel. A sample cartridge further comprising a port). 14. The method of any one of claims 1 to 13, configured to receive a primary output fluid from the primary reaction vessel through a primary output channel fluidly connecting the primary reaction vessel to the secondary reaction vessel. a secondary reaction vessel configured to receive one or more fluid reagents from the reagent vessel through a secondary reagent channel fluidly connecting the reagent vessel to the secondary reaction vessel;
a primary outlet valve disposed in the primary outlet channel to control flow through the primary outlet channel; and
The sample cartridge further comprising a secondary reagent valve disposed in the secondary reagent channel to control flow through the secondary reagent channel. 15. The method of claim 14, wherein the secondary reaction vessel is sealed, and
The sample cartridge is in fluid communication with the secondary reaction vessel and includes a pneumatic module to selectively adjust the pressure of the secondary reaction vessel to draw fluid from the primary outlet channel or secondary reagent channel to the secondary reaction vessel. A sample cartridge further comprising a secondary pneumatic port configured to be connected to. The method of claim 15 , further comprising a secondary pneumatic channel extending between the secondary pneumatic port and the secondary reaction vessel,
The sample cartridge, wherein the opening of the secondary pneumatic channel to the secondary reaction vessel is located somewhat above the side wall of the secondary reaction vessel closer to the top of the secondary reaction vessel than the bottom of the secondary reaction vessel. . 17. The method of any one of claims 14 to 16, wherein the inlet or inlets of the primary output channel and the secondary reagent channel are closer to the top of the secondary reaction vessel than the bottom of the secondary reaction vessel. A sample cartridge that opens upwardly of the side wall of the primary reaction vessel into the secondary reaction vessel. 17. The quality control channel according to any one of claims 13 to 17, wherein at the quality control junction, when relying directly or indirectly on claims 11 or 12, forms an obtuse angle with a portion of the final output channel extending between the quality control joint and the buffer joint. 19. The method according to any one of claims 13 to 18, wherein in the cushioning joint, the cushioning channel has the quality A sample cartridge forming an obtuse angle with a portion of the final output channel extending between a management joint and a buffer joint. 20. The method according to any one of claims 13 to 19, wherein, at the buffer junction, a preset of the final output channel is provided. - a portion of the pre-buffer junction forms an obtuse angle with a portion of the final output channel extending between the quality control junction and the pre-buffer junction,
and wherein, in the quality control junction, a portion of a post-QC junction of the final output channel forms an obtuse angle with a portion of the final output channel extending between the quality control junction and the buffer junction. 1. A sample cartridge for use with a fluid analysis instrument, said cartridge comprising:
a sample vessel configured to receive a fluid sample for analysis;
a buffer container configured to contain a buffer solution;
an assay vessel configured to receive a mixed fluid comprising an aliquot of the fluid sample mixed with at least a portion of the buffer for analysis;
a sample channel extending between the sample vessel and the first junction;
a sample channel valve disposed in the sample channel to control flow of the sample through the sample channel;
a buffer channel extending between the buffer container and the first junction;
a buffer channel valve disposed in the buffer channel to control the flow of the buffer solution through the buffer channel;
a measurement channel in fluid communication with the buffer channel and the sample channel, wherein the measurement channel extends between the first junction and the second junction;
an assay vessel channel in fluid communication with the measurement channel and extending between the second junction and the assay vessel; and
an assay vessel pneumatic port configured to communicate with the assay vessel and be connected to a pneumatic module to selectively adjust the pressure of the assay vessel to draw fluid into the assay vessel through the assay vessel channel, Sample cartridge. 22. The sample cartridge of claim 21, wherein at the first junction, the sample channel forms an obtuse angle with the measurement channel. 23. Sample cartridge according to claim 21 or 22, wherein at the first junction, the buffer channel forms an obtuse angle with the measurement channel. 24. Sample cartridge according to any one of claims 21 to 23, wherein at the second junction the assay vessel channel forms an obtuse angle with the measurement channel. 25. The method of any one of claims 21 to 24, wherein at least one of the sample channel valve and the buffer channel valve is configured to allow an aliquot of the fluid sample to be aspirated into the measurement channel and to allow the buffer to flow through the buffer channel. and an activating valve that can be selectively opened and closed to allow aspiration of the aliquot of the fluid sample for analysis through the measurement channel and the assay vessel channel into the assay vessel. 26. The sample cartridge of any one of claims 21-25, wherein the assay vessel is pre-loaded with a dye configured to mix with the buffer and fluid sample to facilitate analysis. According to any one of claims 21 to 26,
an intermediate outlet in fluid communication with the metering channel through the second junction;
an outlet chamber in which the intermediate outlet is open;
an air-permeable liquid barrier membrane covering the outlet; and
further comprising an intermediate outlet pneumatic port in fluid communication with the outlet chamber and configured to be connected to a pneumatic module to selectively adjust the pressure of the outlet chamber to draw air from the measurement channel through the air-permeable membrane;
and the intermediate outlet is arranged such that liquid entering the measurement channel from the sample channel or buffer channel is allowed to fill the measurement channel, but is not allowed to proceed to the assay vessel channel. 28. The sample cartridge of claim 27, wherein the intermediate outlet is located at the second junction. 28. A second junction and A sample cartridge further comprising an outlet channel extending between the outlets. 30. A sample cartridge according to claim 29, wherein at the second junction, the outlet channel forms an obtuse angle with the measurement channel. According to any one of claims 21 to 30,
an output vessel in fluid communication with the metering channel through the second junction and through an output channel; and
An output vessel pneumatic port in communication with the output vessel and configured to be connected to a pneumatic module to selectively adjust the pressure of the output vessel to draw fluid from the metering channel through the second junction and the output channel to the output vessel. A sample cartridge further comprising: 32. The sample cartridge of claim 31, wherein the output channel extends from the second junction to the output container. 32. A sample cartridge according to claim 31, wherein when according to any one of claims 27 to 30, the output channel extends between the intermediate outlet and the output container. 34. A pressure actuated valve according to any one of claims 21 to 33, wherein the buffer channel valve includes a buffer channel valve pneumatic port configured to be connected to a pneumatic module to selectively open or close the buffer channel valve. A sample cartridge containing an actuated valve. 35. A fluid analysis instrument configured to receive a sample cartridge according to any one of claims 21 to 34, said instrument comprising:
a pneumatic module connected to the assay vessel pneumatic port and configured to selectively adjust the pressure of the assay vessel to draw fluid into the assay vessel through the assay vessel channel; and
An instrument comprising an analysis module configured to measure properties of a fluid in the analysis vessel. 35. A fluid analysis device comprising a sample cartridge according to any one of claims 21 to 34, wherein the device:
a pneumatic module connected to the assay vessel pneumatic port and configured to selectively adjust the pressure of the assay vessel to draw fluid into the assay vessel through the assay vessel channel; and
An instrument comprising an analysis module configured to measure properties of a fluid in the analysis vessel. 37. The method of claim 35 or 36, wherein the analysis module comprises an optical source configured to illuminate the fluid within the analysis vessel, and an optical detector configured to detect or measure light transmitted from the fluid within the analysis vessel. A device containing an optical detector. 38. The method according to any one of claims 35 to 37, when relying directly or indirectly on claim 27, wherein the pneumatic module is further connected or configured to be connected to the intermediate outlet pneumatic port and in the metering channel. A device configured to selectively adjust the pressure of the outlet chamber to draw air through an air-permeable membrane. 39. The method of any one of claims 35 to 38, wherein when relying directly or indirectly on claim 31, the pneumatic module is further connected or configured to be connected to the output vessel pneumatic port and with the second junction. The device is configured to selectively adjust the pressure of the output vessel to draw fluid from the metering channel to the output vessel through the output channel. 39. The method according to any one of claims 35 to 39, when directly or indirectly dependent on claim 65, said pneumatic module is further connected or configured to be connected to said buffer channel valve pneumatic port and said buffer channel valve. A device configured to selectively open or close. 41. The device according to any one of claims 35 or 37 to 40, wherein when relying directly or indirectly on claim 35, the device according to any one of claims 56 to 65 containing fluid samples. An apparatus configured to receive a plurality of sample cartridges. 42. The method of claim 41, wherein a mechanism and an actuator are configured to move the analysis module to various positions corresponding to individual ones of the sample cartridges for analysis of fluid within the analysis vessel of each sample cartridge. A device further comprising: In a fluid analysis system:
The apparatus according to claim 35 or any of claims 37 to 42, when relying directly or indirectly on clause 35,
A fluid analysis system comprising one or more of the sample cartridges of claims 21 to 34. 43. A method of operating a fluid analysis device of claim 36 or any of claims 37 to 42, wherein when relying directly or indirectly on claim 36, said sample vessel contains a fluid sample, said method silver:
operating the pneumatic module to draw sample fluid through the sample channel into a metering channel from the sample vessel to a second junction without the sample fluid advancing into the analysis vessel channel; and
subsequently operating the pneumatic module to aspirate fluid from the buffer vessel through the buffer channel, metering channel and analysis channel to the assay vessel along with an aliquot of the sample fluid from the metering channel. method. According to clause 44,
the pneumatic pressure to reduce the pressure in the assay vessel for a predetermined period of time to draw sample fluid from the sample vessel to the second junction through the sample channel and into the metering channel without the sample fluid proceeding into the assay vessel channel; operating the module; and
to reduce the pressure within the assay vessel after a predetermined period of time for aspirating fluid from the buffer vessel through the buffer channel, metering channel and assay channel with an aliquot of the sample fluid from the metering channel into the assay vessel. The method further comprising operating the pneumatic module. 44. The method of claim 44, when relying directly or indirectly on clause 27, wherein:
operating the pneumatic module to reduce pressure at the intermediate outlet to draw fluid from the buffer container through the sample channel to the metering channel until the sample fluid encounters the air permeable barrier; and
subsequently using the pneumatic pressure to reduce the pressure in the assay vessel to aspirate fluid from the buffer vessel into the assay vessel through the buffer channel, metering channel and analysis channel along with an aliquot of the sample fluid from the metering channel. A method further comprising operating the module. 44. The method of claim 44, when relying directly or indirectly on clause 31, wherein:
Operate the pneumatic module to reduce the pressure in the output vessel for a predetermined period of time to aspirate the sample fluid through the sample channel into the metering channel to a second junction without allowing the sample fluid to advance into the analysis vessel channel. ordering step; and
to reduce the pressure within the assay vessel after a predetermined period of time for aspirating fluid from the buffer vessel through the buffer channel, metering channel and analysis channel along with an aliquot of the sample fluid from the measurement channel into the assay vessel. The method further comprising operating the pneumatic module. 48. The method of any one of claims 44 to 47, wherein operation of the pneumatic module to draw fluid from the buffer container through the buffer channel continues until the metering channel is filled with air. 49. The method of any one of claims 44 to 48, when relying directly or indirectly on clause 31, wherein:
The method further comprising subsequently operating the pneumatic module to reduce the pressure of the output vessel to aspirate sample fluid from the sample vessel into the output vessel. 49. The method of any one of claims 44 to 49, when relying directly or indirectly on clause 34, wherein:
operating the pneumatic module to keep the buffer valve closed during the period during which fluid is aspirated from the sample vessel; and
The method further comprising operating the pneumatic module to hold the buffer valve open so that fluid can then be drawn from the buffer container. 43. A method of operating said fluid analysis device according to any one of claims 35 or 37 to 42, wherein when relying directly or indirectly on claim 35, the fluid in said sample container of said or each sample cartridge Receiving one or more said sample cartridges according to any one of claims 21 to 34 containing a sample, said method comprising:
Operating the pneumatic module to draw the sample fluid from the sample vessel of the or each sample cartridge through the sample channel to the second abutment into the metering channel without advancing into the analysis vessel channel. ;
subsequently operating the pneumatic module to aspirate fluid through the buffer channel, the metering channel and the analysis channel from the buffer container of each sample cartridge into the analysis vessel along with an aliquot of the sample fluid from the metering channel. A method comprising steps. According to clause 51,
Connecting the pneumatic module to the assay vessel pneumatic port of the or each sample cartridge;
within the assay vessel of the or each sample cartridge for a predetermined period of time to aspirate sample fluid from the sample vessel to the second junction through the sample channel and into the metering channel without advancing the sample fluid into the assay vessel channel. operating the pneumatic module to reduce pressure; and
The assay vessel of the or each sample cartridge after the predetermined period of time to aspirate fluid from the buffer vessel through the buffer channel, metering channel and assay channel to the assay vessel along with an aliquot of the sample fluid in the metering channel. The method further comprising operating the pneumatic module to reduce the pressure therein. 51. The method of claim 51, when relying directly or indirectly on clause 59, wherein:
Connecting the pneumatic module to the analysis vessel pneumatic port and the intermediate outlet pneumatic port of the or each sample cartridge;
the pneumatic module to reduce the pressure within the or the intermediate outlet of each sample cartridge to aspirate sample fluid from the sample vessel into the metering channel through the sample channel until the sample fluid meets the air permeable barrier. operating step; and
Fluid is then transferred from the buffer vessel to the assay vessel along with an aliquot of the sample fluid from the metering channel through the buffer channel, metering channel and assay channel to reduce pressure within the assay vessel of the or each sample cartridge. The method further comprising operating the pneumatic module. 51. The method of claim 51, wherein when relying directly or indirectly on clause 31, said method:
Connecting the pneumatic module to the analysis vessel pneumatic port and the output vessel pneumatic port of the or each sample cartridge;
in the output vessel of the or each sample cartridge for a predetermined period of time to aspirate sample fluid from the sample vessel to the second abutment through the sample channel and into the metering channel without advancing the sample fluid into the analysis vessel channel. operating the pneumatic module to reduce pressure; and
within the assay vessel of each sample cartridge after a predetermined period of time to aspirate fluid from the buffer vessel to the assay vessel through the buffer channel, metering channel and assay channel along with an aliquot of the sample fluid in the metering channel. The method further comprising operating the pneumatic module to reduce pressure. 55. The method of any one of claims 51 to 54, wherein operation of the pneumatic module to draw fluid from the buffer container through the buffer channel continues until the metering channel of the or each sample cartridge is filled with air. , method. 56. The method of any one of claims 51 to 55, when relying directly or indirectly on claim 31, wherein:
Connecting the pneumatic module to the output vessel pneumatic port of the or each sample cartridge; and
drawing the buffer into the assay vessel and then operating the pneumatic module to reduce the pressure within the output vessel of the or each sample cartridge to draw sample fluid from the sample vessel into the output vessel. How to. 56. The method of any one of claims 51 to 56, when relying directly or indirectly on clause 34, wherein:
Connecting the pneumatic module to the buffer valve pneumatic port of the or each sample cartridge;
operating the pneumatic module to keep the buffer valve of the or each sample cartridge closed during the period during which fluid is aspirated from the sample vessel; and
The method further comprising subsequently operating the pneumatic module to hold the buffer valve of the or each sample cartridge open such that fluid is drawn from the buffer container. 58. The method of any one of claims 44-57, further comprising subsequently operating the analysis module to measure properties of the fluid within the analysis vessel. 59. The method of claim 58, further comprising transmitting data regarding the measured attribute to an external computing device. A computer-readable storage medium storing instructions that, when executed by a computer, cause the computer to perform the method of any one of claims 44 to 59. A method of using the system of claim 43, said method comprising:
placing a fluid sample into the sample container of the or each sample cartridge;
Inserting the or each sample cartridge into a corresponding cartridge slot of the instrument; and
A method comprising operating the instrument to analyze the fluid sample. 62. The method of claim 61, further comprising removing each sample cartridge from the device once the fluid sample has been processed. In the kit,
A sample cartridge according to any one of claims 7 to 20 if it is directly or indirectly dependent on claim 6 or 6; and
A kit comprising a temporary lid configured to close the output vessel during processing, the temporary lid configured to fluidly connect the final output channel and the output vessel pneumatic channel to the output vessel. 64. The kit of claim 63, wherein the temporary lid is configured to be mechanically coupled to the cartridge by a resiliently flexible body. 65. The kit of claim 64, wherein the body is formed integrally with the temporary lid. 66. A kit according to claim 64 or 65, wherein the body is configured to press the output container against the cartridge when connected. 67. A kit according to any one of claims 64 to 66, wherein the body defines channels for fluidly connecting the final output channel and the output vessel pneumatic channel to the output vessel. 68. The kit of any one of claims 64 to 67, further comprising the output container. A method of using the sample cartridge of any one of claims 1 to 20 or the kit of any of claims 63 to 68, wherein the method comprises extracting, isolating, enriching, concentrating or quantifying or manipulating nucleic acids from a sample; A method comprising operating the device to achieve preparation of nucleic acids for analysis, amplification, sequencing, PCR library preparation, or insertion into a vector. 70. The method of claim 69, wherein the nucleic acid is naturally occurring, non-naturally occurring, DNA, genomic DNA, TCR DNA, cDNA, cfDNA, rearranged immunoglobulin, RNA, mRNA, primary RNA transcript, transfer RNA, microRNA, A method comprising one or more of the following nucleic acid classes: glycolic nucleic acids, threose nucleic acids, locked nucleic acids, and peptide nucleic acids. The method of claim 69 or 70, wherein for said sample:
Processing the sample while maintaining the sample in isolation to prevent contamination of the instrument or cross-contamination with other samples;
Selecting nucleic acids using specific chemistry, incubation conditions, bead selection, and elution parameters;
selecting a desired range of nucleic acid sizes in the processed fluid product and discarding unwanted material outside the desired range;
increasing the concentration of the selected nucleic acid product; and
Performing any two or more of the following processing steps: quantifying an aliquot of the processed fluid product, mixing it with a fluorochrome specific for the selected nucleic acid, and quantifying properties of the product, such as against a standard reference curve. A method further comprising the steps of: