US20210121884A1 - Method of producing a reagent on-board an instrument - Google Patents
Method of producing a reagent on-board an instrument Download PDFInfo
- Publication number
- US20210121884A1 US20210121884A1 US17/066,094 US202017066094A US2021121884A1 US 20210121884 A1 US20210121884 A1 US 20210121884A1 US 202017066094 A US202017066094 A US 202017066094A US 2021121884 A1 US2021121884 A1 US 2021121884A1
- Authority
- US
- United States
- Prior art keywords
- instrument
- concentrate
- diluent
- reagent
- chamber
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 101
- 238000000034 method Methods 0.000 title description 13
- 239000012141 concentrate Substances 0.000 claims abstract description 132
- 239000003085 diluting agent Substances 0.000 claims abstract description 77
- 239000000523 sample Substances 0.000 claims abstract description 46
- 239000002775 capsule Substances 0.000 claims description 26
- 239000012530 fluid Substances 0.000 claims description 13
- 238000004806 packaging method and process Methods 0.000 claims description 13
- 239000012528 membrane Substances 0.000 claims description 12
- 238000007789 sealing Methods 0.000 claims description 12
- 238000011144 upstream manufacturing Methods 0.000 claims description 5
- 239000011261 inert gas Substances 0.000 claims description 3
- 230000000717 retained effect Effects 0.000 claims description 3
- 238000004891 communication Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 238000010186 staining Methods 0.000 description 16
- 238000010790 dilution Methods 0.000 description 10
- 239000012895 dilution Substances 0.000 description 10
- 239000007788 liquid Substances 0.000 description 4
- 230000002378 acidificating effect Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000011888 foil Substances 0.000 description 3
- 238000011532 immunohistochemical staining Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 238000005086 pumping Methods 0.000 description 3
- 238000013207 serial dilution Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- 239000002853 nucleic acid probe Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 1
- 102000052510 DNA-Binding Proteins Human genes 0.000 description 1
- 108700020911 DNA-Binding Proteins Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/52—Containers specially adapted for storing or dispensing a reagent
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/52—Containers specially adapted for storing or dispensing a reagent
- B01L3/523—Containers specially adapted for storing or dispensing a reagent with means for closing or opening
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0605—Metering of fluids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0689—Sealing
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/16—Reagents, handling or storing thereof
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0672—Integrated piercing tool
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/06—Valves, specific forms thereof
- B01L2400/0677—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
- B01L2400/0683—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers mechanically breaking a wall or membrane within a channel or chamber
Definitions
- the present invention relates broadly to producing a reagent on-board an instrument such as a laboratory instrument for treating a biological tissue sample with the reagent.
- the invention also relates generally to an instrument utilising a reagent produced on-board the instrument from a mixture of a concentrate and a diluent.
- Immunohistochemical staining and in situ nucleic acid analysis are tools used in histological diagnosis and the study of tissue morphology. Immunohistochemical staining relies on the specific binding affinity of antibodies with epitopes in tissue samples, and the increasing availability of antibodies which bind specifically with unique epitopes present only in certain types of diseased cellular tissue. Immunohistochemical staining involves a series of treatment steps conducted on a tissue sample (typically a section) mounted on a glass slide to highlight, by selective staining, certain morphological indicators of disease states.
- Typical treatment steps include pretreatment of the tissue sample to reduce non-specific binding, antibody treatment and incubation, enzyme labelled secondary antibody treatment and incubation, substrate reaction with the enzyme to produce a fluorophore or chromophore highlighting areas of the tissue sample having epitopes binding with the antibody, counterstaining, and the like. Between each treatment step, the tissue sample must be rinsed to remove unreacted residual reagent from the prior step. Most treatment steps involve a period of incubation typically conducted at ambient temperature of around 25° C. up to around 40° C., while cell conditioning steps are typically conducted at somewhat higher temperatures, e.g. 90° C. to 100° C.
- In-situ DNA analysis relies upon the specific binding affinity of probes (DNA binding proteins) with unique nucleotide sequences in cell or tissue samples and similarly involves a series of process steps, with a variety of reagents and process temperature requirements. Some specific reactions involve temperatures up to 120° C. to 130° C.
- an instrument utilizing a reagent said instrument providing on-board mixing of the reagent and including:
- the instrument may include one or more mixing wells located on the instrument and configured to receive the concentrate and the diluent for mixing to provide the reagent at the required concentration.
- the mixing well is formed in a sample staining module (SSM) of the instrument.
- SSM sample staining module
- the mixing well may be formed in a fixed or disposable cover member of the SSM. Alternately the mixing well may be a dedicated mixing well formed integral with the instrument.
- the mixing wells are located adjacent one another for serial dilution of the concentrate with the diluent.
- the chamber may be a container supplied in a sealed condition and adapted to contain a predetermined volume of the concentrate.
- the sealed container may include a sealing membrane arranged to contain the concentrate and an inert gas.
- the sealing membrane may be designed to be at least partly removed or pierced and the container is sized for addition of diluent for mixing on-board the instrument to provide the reagent at the required concentration.
- the chamber comprises a plurality of the sealed containers each adapted to contain a predetermined volume of the concentrate.
- the plurality of chambers are arranged as multiple reservoirs e.g. in a blister pack.
- the chamber is a disposable or reusable probe tip containing the concentrate and adapted to fit to a dispensing probe of the instrument.
- the instrument includes a concentrate dispenser for on-board provision of a reagent, said dispenser comprising packaging adapted to contain a plurality of concentrate capsules, the packaging configured to release each of the capsules for mixing with a diluent on-board the instrument to provide the reagent at the required concentration.
- the packaging includes a stacked cartridge adapted to contain one or more columns of the plurality of concentrate capsules. In some embodiments two or more columns of the concentrate capsules are provided in the cartridge. More preferably the dispenser also comprises a reciprocating release operatively coupled to the stacked cartridge to release a lowermost of the column of capsules form the cartridge for mixing with the diluent on-board the instrument.
- the packaging includes a cartridge reel adapted to contain a roll of the concentrate capsules retained in a concentrate tape configured to be dispensed from the cartridge reel. In this embodiment the concentrate tape includes two strips adhered to one another with the concentrate capsules sandwiched therebetween and adapted to be released from the concentrate tape.
- an instrument utilising a reagent said instrument including:
- the diluent chamber may be a vacuum chamber to which the concentrate dispenser is operatively coupled wherein the concentrate is dispensed into the vacuum chamber with the assistance of vacuum pressure applied to the diluent chamber.
- the concentrate dispenser may include a plunger for dispensing of the concentrate into the diluent chamber, preferably via a non-return valve.
- the concentrate dispenser may include a frangible separator which cooperates with the diluent chamber wherein rupturing of the frangible separator effects dispensing of the concentrate to the diluent chamber.
- the instrument is a laboratory instrument and the reagent is a high value reagent such as a molecular probe such as a nucleic acid probe or an antibody (Ab) although that need not be the case and the reagent may be a bulk reagent such as a wash fluid or buffer which is formulated on-board the instrument from concentrate and diluent.
- the concentrate may be in any state such as a liquid, solid, or lyophilised state.
- Also described herein is a method of producing a reagent on-board an instrument, said method including the steps of:
- the method may also comprise one or more subsequent steps of diluting the reagent with additional diluent to produce the required concentration.
- packaging for use with an instrument in the on-board provision of a reagent, the packaging being adapted to contain a plurality of concentrate capsules, and configured to release each of the capsules for mixing with a diluent on-board the instrument to provide the reagent.
- a container for use with an instrument in the on-board provision of a reagent the container adapted to contain a predetermined volume of a concentrate for mixing with a diluent on-board the instrument to provide the reagent.
- cover member for use with an instrument in the on-board provision of a reagent, the cover member including a well being adapted to receive a concentrate for mixing with a diluent on-board the instrument to provide the reagent.
- FIG. 1A is a schematic illustration of a cover member associated with a sample staining module (SSM) of an instrument having on-board production of the reagent according to an embodiment of one aspect of the invention
- FIG. 1B and FIG. 1C show a concentrate packaged in a sealed container for dispensing into the cover member of FIG. 1A
- FIG. 1A , FIG. 1B , and FIG. 1C may be collectively considered as a FIG. 1 .
- FIG. 2A and FIG. 2B are schematics of alternative embodiments of mixing wells or chambers designed for on-board production of reagents; FIG. 2A and FIG. 2B may be collectively considered as a FIG. 2 .
- FIG. 3A and FIG. 3B are schematics of alternative embodiments of chambers designed to be used in conjunction with the instrument described with respect to FIG. 1A ;
- FIG. 3A and FIG. 3B show side and top views respectively;
- FIG. 3A and FIG. 3B may be collectively considered as a FIG. 3 .
- FIG. 4A , FIG. 4B , FIG. 4C , and FIG. 4D are schematics of different concentrate dispensers designed for the on-board provision of a reagent at an instrument according to embodiments of another aspect of the invention; FIG. 4A .
- FIG. 4B , FIG. 4C , and FIG. 4D may be collectively considered as a FIG. 4 .
- FIG. 5A , FIG. 5B , and FIG. 5C are schematics of packaged concentrates/diluents used to produce reagents on-board an instrument according to embodiments of a further aspect of the invention;
- FIG. 5A shows the sealing membrane in-tact (left) and ruptured (right);
- FIG. 5 a , FIG. 5B , and FIG. 5C may be collectively considered as a FIG. 5 .
- FIG. 6 is a schematic illustration of an instrument utilising a reagent and providing on-board mixing of the reagent according to embodiments of the invention.
- FIG. 7 is a flow diagram showing steps in a method of producing reagent onboard an instrument according to embodiments of the invention.
- FIG. 1A is a schematic illustration of a cover member 14 of a slide staining module (SSM) 74 of an instrument 70 ( FIG. 6 ).
- well 10 may be utilised as a mixing well for mixing a concentrate 12 with a diluent (not shown) on-board the instrument 70 to provide a reagent at a required concentration.
- the well 10 is in this embodiment formed in a cover member 14 associated with a sample staining module (SSM) 74 of the instrument 70 and may receive or mix reagent.
- the cover member 14 may be releasably attachable to the SSM 74 of the instrument 70 .
- the mixing well in alternative embodiments may be formed as an integral part of the instrument.
- instrument 70 is provided with multiple mixing wells for serial dilution of the concentrate as illustrated in FIG. 2A .
- the mixing well is a mixing chamber which is integral with a reagent dispensing probe 30 of the instrument, as illustrated in FIG. 2B .
- the instrument 70 of this aspect of the invention is a laboratory instrument for treating a biological tissue sample within the SSM with the reagent, typically a high value reagent in the form of a molecular probe such as a nucleic acid probe or an antibody (Ab).
- the high value reagent is produced at a desired concentration on-board the instrument 70 by mixing the concentrate such as 12 and diluent to provide the reagent at the required concentration.
- the reagent need not be “high value” and may be any concentrate that is amenable to dilution to a required concentration such as, for example, wash fluid or buffer solution which may be diluted with for example, deionised water, to a predetermined concentration suitable for e.g. treatment of the sample or washing or other use within a cover member of the SSM.
- FIG. 6 is a schematic illustration of an instrument 70 according to some embodiments of the invention, including a plurality of side staining modules 74 arranged to receive slides 73 .
- At least one slide transport robot 78 in the form of a gantry robot configured by a controller (not shown) moves the slides 73 using a slide transport device 75 (such as a suction cup) to and from the slide staining modules 74 .
- the fluid transport system 72 includes a fluid dispensing robot 76 configured by the controller to dispense a plurality of reagents from a probe 30 of the fluid dispensing robot to the slides 73 received in the slide treatment modules 74 to treat the tissue samples on the slides.
- a designated combination of reagents e.g. higher and lesser value reagents, are required to be dispensed to a slide and in some embodiments it is desirable to achieve mixing to a predetermined concentration on-board the instrument.
- the fluid dispensing robot 76 is configured by the controller to dispense a plurality of reagents stored in reagent containers 80 , 81 on board the instrument to the slides 73 in the slide staining modules 74 .
- the concentrate 12 is provided in a sealed container 13 .
- the sealed container 13 includes a sealing membrane 18 arranged to contain the concentrate 12 within a plurality of sealed chambers 16 .
- the sealed container 13 of this example is in the form of a blister pack 20 .
- the blister pack 20 is designed to release each of the concentrate volumes such as 12 into a chamber such as a concentrate chamber or a mixing chamber for on-board mixing of the reagent.
- each of the concentrate volumes such as 12 is released into the well 10 of the cover member 14 .
- the sealing membrane 18 which is in the form of a frangible foil is ruptured or otherwise broken or removed by the instrument across an entry to the well 10 (or instrument chamber) for release of the concentrate 12 .
- the diluent may be pumped from a bulk supply into the well 10 either before or after addition of the concentrate to achieve the required concentration.
- the reagent is thus mixed and produced on-board the instrument for dispensing e.g. by a probe to cover member 14 .
- the reagent may then be drawn under the cover member 14 or aspirated using a robot and a fluid transport probe (not shown) and then dispensed through a port 9 into a reaction area 11 for treatment, such as staining.
- FIG. 2A and FIG. 2B illustrate alternative embodiments suitable for production of a reagent on-board an instrument such as that including the cover member 14 of the previous embodiment of FIG. 1A .
- the same reference numerals have been used for similar components of this and any other embodiments.
- the instrument 70 is provided with multiple mixing wells 10 A to 10 D for serial dilution of the concentrate 12 .
- the reagent 12 is at stage 1 transferred to the mixing well 10 B and at stage 2 diluted with diluent.
- the resulting reagent is mixed in the mixing well 10 B and at stage 3 transferred to mixing well 10 C for further dilution at stage 4 with diluent.
- the resulting reagent is mixed in mixing well 10 C and at stage 5 transferred to mixing well 10 D for further dilution with diluent at stage 6 .
- the final reagent at its required concentration is at stage 7 transferred from the mixing well 10 D for use by the instrument 70 .
- the number of mixing wells required for each desired dilution may vary according to the required dilution. Accordingly, the number of mixing wells provided or utilized may vary according to the required dilution.
- the final reagent may for example be used in staining a tissue sample located on a slide (not shown) of the SSM 74 .
- the multiple mixing wells 10 A to 10 D may be formed as an integral part of the instrument 70 in for example the SSM 74 or as shown in FIG. 1A fabricated in a fixed or disposable cover member of the SSM.
- the instrument 70 includes a probe 30 having an in-line mixing chamber 32 to provide on-board production of a reagent.
- the probe 30 is for example in the form of a fluid transfer probe (FTP) of an instrument used for dispensing fluids for treating or staining tissue samples on slides in the SSM 74 .
- FTP fluid transfer probe
- the modified FTP 30 includes a bifurcation to a diluent chamber 34 and a concentrate chamber 36 located up stream of the mixing chamber 32 .
- the diluent and concentrate chambers 34 / 36 provide the requisite volume of diluent 38 and concentrate 40 to the mixing chamber 32 to provide a reagent at the required concentration.
- the FTP 30 includes a nozzle 42 downstream of the mixing chamber 32 which is configured to dispense the mixed reagent into a cover member 14 of a SSM 74 .
- the nozzle 42 may discharge reagent directly on to the sample slide or into a port or well of a cover member 14 (see FIG. 1A ) associated with the SSM.
- the instrument may incorporate a separate valve or valves (not shown) arranged on or adjacent to the mixing chamber 32 .
- the mixing chamber 32 may be in fluid communication with further concentrate and diluent reservoirs ( 80 , 81 ) and optionally, further concentrate and diluent chambers ( 36 , 34 ) upstream of the mixing chamber 32 such that the probe receives more than two constituents for mixing, enabling flexibility in the combinations and concentrations of reagents that are prepared on-board the instrument 70 as may be determined by a controller of the instrument.
- the concentrate reservoirs 80 may be e.g. in the form of sealed containers as disclosed herein.
- FIG. 7 is a flow chart illustrating steps in a method of producing a reagent on-board the instrument.
- the method includes, in a step 701 , pumping a requisite volume of concentrate into the concentrate chamber and, in a step 702 , pumping a requisite volume of diluent into the diluent chamber to achieve a required concentrate of reagent.
- steps 703 and 704 the concentrate is released from the concentrate chamber and the diluent chamber respectively into the mixing chamber in the probe.
- the reagent is mixed at the required concentration
- the mixed reagent is dispensed from the probe to the cover member.
- the instrument controller may be configured to implement the preferred method of on-board mixing of reagents by controlling operation of the fluid transport system 72 including the fluid dispensing robot 76 and pumping of concentrate 40 and diluent 38 from containers 80 , 81 respectively, into the mixing chamber 32 either directly, or through concentrate and diluent chambers 36 , 34 located in the probe 30 upstream of the mixing chamber as is shown in FIG. 7 .
- FIG. 7 also shows various instrument features 80 , 20 , 50 and 81 , for storing and delivering concentrate and diluent for mixing of reagent on board the instrument.
- FIG. 3A and FIG. 3B are alternate embodiments and views of a chamber or concentrate container suitable for use with an instrument providing on-board production of a reagent.
- the chambers may for example be suitable for use with the cover member 14 of the instrument of FIG. 1A , or to supply the mixing wells or mixing chambers of FIG. 2A and 2B .
- the concentrate such as 12 is contained in a chamber 16 which is sealed with a sealing membrane 18 .
- the concentrate 12 is protected from evaporation and oxidation by the use of nitrogen or other inert gases 22 sealed within the chamber 16 .
- the sealing membrane 18 may be in the form of a foil pack or lid seal.
- the embodiment of FIG. 3B is similar to the blister pack 20 of FIG. 1B having multiple chambers such as 16 in a common base plate 24 . Although not shown the multiple chambers 16 may also be sealed with a common sealing membrane or foil.
- FIG. 4A , FIG. 4B , FIG. 4C , and FIG. 4D illustrate different embodiments of a concentrate dispenser for use on-board an instrument in production of a reagent, for example the reagent being used in staining or otherwise treating tissue samples in an SSM of the instrument.
- the embodiment of FIG. 4A schematically depicts mixing of a concentrate 12 with a diluent 13 to provide the reagent as may be achieved using mixing wells and mixing chambers on-board the instrument, according to various embodiments disclosed herein.
- the concentrate 12 is in this embodiment is contained in a dissolving capsule 15 designed to dissolve during mixing with the diluent 13 .
- the diluent 13 may be acidic or basic depending on the chemistry of the concentrate, and the capsule 15 formed of a material which dissolves in the diluent 13 .
- the diluent 13 may be contained in a mixing well associated with a cover member 14 of the SSM in FIG. 1A , or a mixing well, diluent chamber or mixing chamber of the instrument as illustrated in FIG. 2A and FIG. 2B .
- the concentrate such as 12 is contained in a concentrate dispenser 50 operatively coupled to an instrument for on-board provision of a reagent.
- the concentrate dispenser 50 includes packaging 52 within which one of a plurality of the concentrate capsules such as 12 A to 12 F are contained in a stacked cartridge.
- the stacked cartridge packaging 52 is designed to release each of the capsules such as 12 A for mixing with a diluent on-board the instrument to provide a reagent at the required concentration.
- the concentrate dispenser 50 includes a reciprocating release 54 operatively coupled to the stacked cartridge 52 to release a lowermost of the column of capsules such as 12 A.
- the packaging of the concentrate dispenser 50 is in the form of a cartridge reel 52 contained within the instrument.
- the cartridge reel contains a roll of the concentrate capsules such as 12 A to 12 F retained in a concentrate tape 56 dispensed from the cartridge reel 52 .
- the concentrate tape 56 includes two strips 58 A and 58 B adhered to one another with the concentrate capsules such as 12 F sandwiched therebetween.
- the concentrate dispenser 50 of FIG. 4C also includes a spool 60 within the instrument for removing the lower strip 58 B of the concentrate tape 56 to release each of the concentrate capsules such as 12 A from the dispenser 50 .
- the instrument includes a chamber 16 in the form of a disposable or reusable probe tip containing the concentrate 12 .
- the probe tip 16 is designed to fit to a probe such as 44 of the concentrate dispenser.
- the probe may for example be in the form of a FTP of an instrument used in treating or staining tissue samples.
- the probe tip 16 includes a nozzle 46 designed to cooperate with a port of for example a cover member of a SSM.
- the nozzle 46 includes a temporary seal 48 which is designed to rupture on discharge of the concentrate 12 into the well for mixing with the diluent.
- the concentrate 12 may also be sealed in the probe tip 16 with another seal 49 which is similarly ruptured under the application of pressure applied by the FTP 44 .
- FIG. 5A , FIG. 5B , and FIG. 5C illustrate various embodiments of a diluent chamber 17 operatively coupled to a concentrate dispenser 19 for mixing on-board an instrument to produce a reagent.
- the diluent chamber is in the form of a mixing vial 17 with the concentrate dispenser 19 attached as a single package.
- the diluent vial 17 has a vacuum pressure applied to it for dispensing of the concentrate 12 from the concentrate dispenser 19 .
- the concentrate dispenser 19 includes a plunger 21 for urging the concentrate 12 from the concentrate dispenser 19 into the diluent vial 17 .
- the concentrate dispenser 19 may include a one way valve 23 through which the concentrate 12 is discharged into the diluent vial 17 .
- the embodiment of FIG. 5C includes a concentrate dispenser in the form of the packaged chamber 16 such as the embodiments of FIG. 3A .
- the chamber 16 is operatively coupled to the diluent chamber or vial 17 with its sealing membrane 18 in contact with an entrance 25 to the diluent chamber 17 .
- the entrance 25 includes a spiked ridge 27 designed on the application of downward pressure to the chamber 16 to rupture the frangible sealing membrane 18 .
- the concentrate 12 is thus discharged into the diluent chamber 17 for mixing to produce the necessary reagent.
- a concentrate includes liquids of varying concentrations and for example includes high-value reagents and bulk reagents.
- the FTP of the earlier embodiment may thus extend to dispensing a reagent to be diluted, a diluent, other reagents, or a combination of these liquids.
- the instrument includes a controller configured to implement the preferred method of producing a reagent on-board the instrument.
- the method may be implemented via computer program code including instructions to control movement of a concentrate dispenser in, for example, dispensing a concentrate to a well of a cover member associated with a SSM as in FIG. 1A , or a mixing well, diluent chamber or mixing chamber of the instrument as illustrated in FIGS. 2A and 2B .
- the controller may also control dilution and mixing of the concentrate with a diluent to provide the reagent at the required concentration.
- the concentration of the reagent produced on-board the instrument can be varied, for example, to achieve the required staining option for tissue samples;
- On-board dilution improves laboratory workflow without having to pre-dilute reagents from concentrates and also reduces operator time and potential errors.
Abstract
The present invention relates broadly to production of a reagent on-board an instrument. The instrument is provided with a probe for dispensing said reagent, a concentrate chamber adapted to contain a concentrate and a diluent chamber adapted to contain a diluent. The probe includes an in-line mixing chamber adapted to receive the concentrate and the diluent to provide the reagent at the required concentration for dispensing by the probe.
Description
- This application is a continuation-in-part of U.S. application Ser. No. 14/653,956 filed on Jun. 19, 2015 which is a National Stage of International Application No. PCT/AU2013/001499 filed Dec. 20, 2013, claiming priority based on U.S. Provisional Application No. 61/745,331 filed Dec. 21, 2012, the contents of all of which are incorporated herein by reference in their entirety.
- The present invention relates broadly to producing a reagent on-board an instrument such as a laboratory instrument for treating a biological tissue sample with the reagent. The invention also relates generally to an instrument utilising a reagent produced on-board the instrument from a mixture of a concentrate and a diluent.
- Immunohistochemical staining and in situ nucleic acid analysis are tools used in histological diagnosis and the study of tissue morphology. Immunohistochemical staining relies on the specific binding affinity of antibodies with epitopes in tissue samples, and the increasing availability of antibodies which bind specifically with unique epitopes present only in certain types of diseased cellular tissue. Immunohistochemical staining involves a series of treatment steps conducted on a tissue sample (typically a section) mounted on a glass slide to highlight, by selective staining, certain morphological indicators of disease states.
- Typical treatment steps include pretreatment of the tissue sample to reduce non-specific binding, antibody treatment and incubation, enzyme labelled secondary antibody treatment and incubation, substrate reaction with the enzyme to produce a fluorophore or chromophore highlighting areas of the tissue sample having epitopes binding with the antibody, counterstaining, and the like. Between each treatment step, the tissue sample must be rinsed to remove unreacted residual reagent from the prior step. Most treatment steps involve a period of incubation typically conducted at ambient temperature of around 25° C. up to around 40° C., while cell conditioning steps are typically conducted at somewhat higher temperatures, e.g. 90° C. to 100° C. In-situ DNA analysis relies upon the specific binding affinity of probes (DNA binding proteins) with unique nucleotide sequences in cell or tissue samples and similarly involves a series of process steps, with a variety of reagents and process temperature requirements. Some specific reactions involve temperatures up to 120° C. to 130° C.
- In these various treatment steps it is understood that no single concentration of antibody is sufficient to account for tissue variations, processing variability and tissue sample thickness. Reagents are provided in a Ready to Use (RTU) format that cannot be varied in concentration. Alternatively, concentrates are supplied and diluted to achieve a range of staining options. However, these approaches do not address increasing customer need for workflow flexibility with ease of reagent uses.
- According to a first aspect of the invention there is provided an instrument utilizing a reagent, said instrument providing on-board mixing of the reagent and including:
-
- a probe for dispensing said reagent;
- a concentrate chamber adapted to contain a concentrate; and
- a diluent chamber adapted to contain a diluent;
- wherein the probe includes an in-line mixing chamber adapted to receive the concentrate and the diluent to provide the reagent at the required concentration for dispensing by the probe.
- The instrument may include one or more mixing wells located on the instrument and configured to receive the concentrate and the diluent for mixing to provide the reagent at the required concentration. In one embodiment the mixing well is formed in a sample staining module (SSM) of the instrument. The mixing well may be formed in a fixed or disposable cover member of the SSM. Alternately the mixing well may be a dedicated mixing well formed integral with the instrument.
- Preferably the mixing wells are located adjacent one another for serial dilution of the concentrate with the diluent.
- The chamber may be a container supplied in a sealed condition and adapted to contain a predetermined volume of the concentrate. The sealed container may include a sealing membrane arranged to contain the concentrate and an inert gas. The sealing membrane may be designed to be at least partly removed or pierced and the container is sized for addition of diluent for mixing on-board the instrument to provide the reagent at the required concentration. Alternatively or additionally the chamber comprises a plurality of the sealed containers each adapted to contain a predetermined volume of the concentrate. In this embodiment the plurality of chambers are arranged as multiple reservoirs e.g. in a blister pack. In another embodiment the chamber is a disposable or reusable probe tip containing the concentrate and adapted to fit to a dispensing probe of the instrument.
- therein some embodiments, the instrument includes a concentrate dispenser for on-board provision of a reagent, said dispenser comprising packaging adapted to contain a plurality of concentrate capsules, the packaging configured to release each of the capsules for mixing with a diluent on-board the instrument to provide the reagent at the required concentration.
- Preferably the packaging includes a stacked cartridge adapted to contain one or more columns of the plurality of concentrate capsules. In some embodiments two or more columns of the concentrate capsules are provided in the cartridge. More preferably the dispenser also comprises a reciprocating release operatively coupled to the stacked cartridge to release a lowermost of the column of capsules form the cartridge for mixing with the diluent on-board the instrument. Alternatively the packaging includes a cartridge reel adapted to contain a roll of the concentrate capsules retained in a concentrate tape configured to be dispensed from the cartridge reel. In this embodiment the concentrate tape includes two strips adhered to one another with the concentrate capsules sandwiched therebetween and adapted to be released from the concentrate tape.
- Also described herein is an instrument utilising a reagent, said instrument including:
-
- a diluent chamber adapted to contain a diluent;
- a concentrate dispenser operatively coupled to the diluent chamber to dispense concentrate for mixing with the diluent on-board the instrument to produce the reagent at a required concentration. the concentrate may be contained in dissolvable capsules designed to dissolve during mixing with the diluent to produce the reagent. In this embodiment the diluent is acidic or basic depending on the concentrate, and the dissolvable capsules are formed at least in part of a material which dissolves in the acidic or basic diluent.
- The diluent chamber may be a vacuum chamber to which the concentrate dispenser is operatively coupled wherein the concentrate is dispensed into the vacuum chamber with the assistance of vacuum pressure applied to the diluent chamber. Alternatively the concentrate dispenser may include a plunger for dispensing of the concentrate into the diluent chamber, preferably via a non-return valve. Still alternatively the concentrate dispenser may include a frangible separator which cooperates with the diluent chamber wherein rupturing of the frangible separator effects dispensing of the concentrate to the diluent chamber.
- Generally the instrument is a laboratory instrument and the reagent is a high value reagent such as a molecular probe such as a nucleic acid probe or an antibody (Ab) although that need not be the case and the reagent may be a bulk reagent such as a wash fluid or buffer which is formulated on-board the instrument from concentrate and diluent. The concentrate may be in any state such as a liquid, solid, or lyophilised state.
- Also described herein is a method of producing a reagent on-board an instrument, said method including the steps of:
-
- providing a concentrate at the instrument;
- discharging the concentrate into a diluent provided at the instrument, or vice versa;
- mixing the concentrate with the diluent on-board the instrument to provide the reagent at a required concentration.
- In one embodiment the method may also comprise one or more subsequent steps of diluting the reagent with additional diluent to produce the required concentration.
- Also described is packaging for use with an instrument in the on-board provision of a reagent, the packaging being adapted to contain a plurality of concentrate capsules, and configured to release each of the capsules for mixing with a diluent on-board the instrument to provide the reagent.
- Also described is a container for use with an instrument in the on-board provision of a reagent, the container adapted to contain a predetermined volume of a concentrate for mixing with a diluent on-board the instrument to provide the reagent.
- Also described is a cover member for use with an instrument in the on-board provision of a reagent, the cover member including a well being adapted to receive a concentrate for mixing with a diluent on-board the instrument to provide the reagent.
- In order to achieve a better understanding of the nature of the present invention a preferred embodiment of an instrument utilizing a reagent will now be described, by way of example only, with reference to the accompanying drawings in which:
-
FIG. 1A is a schematic illustration of a cover member associated with a sample staining module (SSM) of an instrument having on-board production of the reagent according to an embodiment of one aspect of the invention;FIG. 1B andFIG. 1C show a concentrate packaged in a sealed container for dispensing into the cover member ofFIG. 1A ;FIG. 1A ,FIG. 1B , andFIG. 1C may be collectively considered as aFIG. 1 . -
FIG. 2A andFIG. 2B are schematics of alternative embodiments of mixing wells or chambers designed for on-board production of reagents;FIG. 2A andFIG. 2B may be collectively considered as aFIG. 2 . -
FIG. 3A andFIG. 3B are schematics of alternative embodiments of chambers designed to be used in conjunction with the instrument described with respect toFIG. 1A ;FIG. 3A andFIG. 3B show side and top views respectively;FIG. 3A andFIG. 3B may be collectively considered as aFIG. 3 . -
FIG. 4A ,FIG. 4B ,FIG. 4C , andFIG. 4D are schematics of different concentrate dispensers designed for the on-board provision of a reagent at an instrument according to embodiments of another aspect of the invention;FIG. 4A .FIG. 4B ,FIG. 4C , andFIG. 4D may be collectively considered as aFIG. 4 . -
FIG. 5A ,FIG. 5B , andFIG. 5C are schematics of packaged concentrates/diluents used to produce reagents on-board an instrument according to embodiments of a further aspect of the invention;FIG. 5A shows the sealing membrane in-tact (left) and ruptured (right);FIG. 5a ,FIG. 5B , andFIG. 5C may be collectively considered as aFIG. 5 . -
FIG. 6 is a schematic illustration of an instrument utilising a reagent and providing on-board mixing of the reagent according to embodiments of the invention. -
FIG. 7 is a flow diagram showing steps in a method of producing reagent onboard an instrument according to embodiments of the invention. -
FIG. 1A is a schematic illustration of acover member 14 of a slide staining module (SSM) 74 of an instrument 70 (FIG. 6 ). In one embodiment, well 10 may be utilised as a mixing well for mixing aconcentrate 12 with a diluent (not shown) on-board theinstrument 70 to provide a reagent at a required concentration. The well 10 is in this embodiment formed in acover member 14 associated with a sample staining module (SSM) 74 of theinstrument 70 and may receive or mix reagent. Thecover member 14 may be releasably attachable to theSSM 74 of theinstrument 70. The mixing well in alternative embodiments may be formed as an integral part of the instrument. For example, in some embodiments,instrument 70 is provided with multiple mixing wells for serial dilution of the concentrate as illustrated inFIG. 2A . In other embodiments, the mixing well is a mixing chamber which is integral with areagent dispensing probe 30 of the instrument, as illustrated inFIG. 2B . - The
instrument 70 of this aspect of the invention is a laboratory instrument for treating a biological tissue sample within the SSM with the reagent, typically a high value reagent in the form of a molecular probe such as a nucleic acid probe or an antibody (Ab). The high value reagent is produced at a desired concentration on-board theinstrument 70 by mixing the concentrate such as 12 and diluent to provide the reagent at the required concentration. It is to be understood, however, that the reagent need not be “high value” and may be any concentrate that is amenable to dilution to a required concentration such as, for example, wash fluid or buffer solution which may be diluted with for example, deionised water, to a predetermined concentration suitable for e.g. treatment of the sample or washing or other use within a cover member of the SSM. -
FIG. 6 is a schematic illustration of aninstrument 70 according to some embodiments of the invention, including a plurality ofside staining modules 74 arranged to receive slides 73. At least oneslide transport robot 78 in the form of a gantry robot configured by a controller (not shown) moves theslides 73 using a slide transport device 75 (such as a suction cup) to and from theslide staining modules 74. Thefluid transport system 72 includes afluid dispensing robot 76 configured by the controller to dispense a plurality of reagents from aprobe 30 of the fluid dispensing robot to theslides 73 received in theslide treatment modules 74 to treat the tissue samples on the slides. - In some cases, to treat the tissue samples on the
slides 73, a designated combination of reagents, e.g. higher and lesser value reagents, are required to be dispensed to a slide and in some embodiments it is desirable to achieve mixing to a predetermined concentration on-board the instrument. Thefluid dispensing robot 76 is configured by the controller to dispense a plurality of reagents stored inreagent containers slides 73 in theslide staining modules 74. - In the embodiment of
FIG. 1B andFIG. 1C theconcentrate 12 is provided in a sealedcontainer 13. The sealedcontainer 13 includes a sealingmembrane 18 arranged to contain theconcentrate 12 within a plurality of sealedchambers 16. The sealedcontainer 13 of this example is in the form of ablister pack 20. Theblister pack 20 is designed to release each of the concentrate volumes such as 12 into a chamber such as a concentrate chamber or a mixing chamber for on-board mixing of the reagent. In the context ofFIG. 1A , each of the concentrate volumes such as 12 is released into the well 10 of thecover member 14. The sealingmembrane 18 which is in the form of a frangible foil is ruptured or otherwise broken or removed by the instrument across an entry to the well 10 (or instrument chamber) for release of theconcentrate 12. In this and other embodiments, the diluent may be pumped from a bulk supply into the well 10 either before or after addition of the concentrate to achieve the required concentration. The reagent is thus mixed and produced on-board the instrument for dispensing e.g. by a probe to covermember 14. The reagent may then be drawn under thecover member 14 or aspirated using a robot and a fluid transport probe (not shown) and then dispensed through aport 9 into areaction area 11 for treatment, such as staining. -
FIG. 2A andFIG. 2B illustrate alternative embodiments suitable for production of a reagent on-board an instrument such as that including thecover member 14 of the previous embodiment ofFIG. 1A . For ease of reference and to avoid repetition, the same reference numerals have been used for similar components of this and any other embodiments. In the embodiment ofFIG. 2A theinstrument 70 is provided withmultiple mixing wells 10A to 10D for serial dilution of theconcentrate 12. Thereagent 12 is at stage 1 transferred to the mixing well 10B and atstage 2 diluted with diluent. The resulting reagent is mixed in the mixing well 10B and atstage 3 transferred to mixing well 10C for further dilution atstage 4 with diluent. The resulting reagent is mixed in mixing well 10C and atstage 5 transferred to mixing well 10D for further dilution with diluent atstage 6. The final reagent at its required concentration is atstage 7 transferred from themixing well 10D for use by theinstrument 70. It will be appreciated by those skilled in the art that the number of mixing wells required for each desired dilution may vary according to the required dilution. Accordingly, the number of mixing wells provided or utilized may vary according to the required dilution. The final reagent may for example be used in staining a tissue sample located on a slide (not shown) of theSSM 74. Themultiple mixing wells 10A to 10D may be formed as an integral part of theinstrument 70 in for example theSSM 74 or as shown inFIG. 1A fabricated in a fixed or disposable cover member of the SSM. - In the embodiment of
FIG. 2B theinstrument 70 includes aprobe 30 having an in-line mixing chamber 32 to provide on-board production of a reagent. Theprobe 30 is for example in the form of a fluid transfer probe (FTP) of an instrument used for dispensing fluids for treating or staining tissue samples on slides in theSSM 74. The modifiedFTP 30 includes a bifurcation to adiluent chamber 34 and aconcentrate chamber 36 located up stream of the mixingchamber 32. The diluent and concentratechambers 34/36 provide the requisite volume ofdiluent 38 and concentrate 40 to the mixingchamber 32 to provide a reagent at the required concentration. TheFTP 30 includes anozzle 42 downstream of the mixingchamber 32 which is configured to dispense the mixed reagent into acover member 14 of aSSM 74. Thenozzle 42 may discharge reagent directly on to the sample slide or into a port or well of a cover member 14 (seeFIG. 1A ) associated with the SSM. In one embodiment the instrument may incorporate a separate valve or valves (not shown) arranged on or adjacent to the mixingchamber 32. It is to be understood that the mixingchamber 32 may be in fluid communication with further concentrate and diluent reservoirs (80, 81) and optionally, further concentrate and diluent chambers (36, 34) upstream of the mixingchamber 32 such that the probe receives more than two constituents for mixing, enabling flexibility in the combinations and concentrations of reagents that are prepared on-board theinstrument 70 as may be determined by a controller of the instrument. Theconcentrate reservoirs 80 may be e.g. in the form of sealed containers as disclosed herein. -
FIG. 7 is a flow chart illustrating steps in a method of producing a reagent on-board the instrument. The method includes, in astep 701, pumping a requisite volume of concentrate into the concentrate chamber and, in astep 702, pumping a requisite volume of diluent into the diluent chamber to achieve a required concentrate of reagent. Insteps step 705, the reagent is mixed at the required concentration, and in astep 706, the mixed reagent is dispensed from the probe to the cover member. It is to be understood that the instrument controller may be configured to implement the preferred method of on-board mixing of reagents by controlling operation of thefluid transport system 72 including thefluid dispensing robot 76 and pumping ofconcentrate 40 and diluent 38 fromcontainers chamber 32 either directly, or through concentrate anddiluent chambers probe 30 upstream of the mixing chamber as is shown inFIG. 7 . -
FIG. 7 also shows various instrument features 80, 20, 50 and 81, for storing and delivering concentrate and diluent for mixing of reagent on board the instrument. -
FIG. 3A andFIG. 3B are alternate embodiments and views of a chamber or concentrate container suitable for use with an instrument providing on-board production of a reagent. The chambers may for example be suitable for use with thecover member 14 of the instrument ofFIG. 1A , or to supply the mixing wells or mixing chambers ofFIG. 2A and 2B . In the various embodiments ofFIG. 3A the concentrate such as 12 is contained in achamber 16 which is sealed with a sealingmembrane 18. Theconcentrate 12 is protected from evaporation and oxidation by the use of nitrogen or otherinert gases 22 sealed within thechamber 16. The sealingmembrane 18 may be in the form of a foil pack or lid seal. The embodiment ofFIG. 3B is similar to theblister pack 20 ofFIG. 1B having multiple chambers such as 16 in acommon base plate 24. Although not shown themultiple chambers 16 may also be sealed with a common sealing membrane or foil. -
FIG. 4A ,FIG. 4B ,FIG. 4C , andFIG. 4D illustrate different embodiments of a concentrate dispenser for use on-board an instrument in production of a reagent, for example the reagent being used in staining or otherwise treating tissue samples in an SSM of the instrument. The embodiment ofFIG. 4A schematically depicts mixing of aconcentrate 12 with a diluent 13 to provide the reagent as may be achieved using mixing wells and mixing chambers on-board the instrument, according to various embodiments disclosed herein. Theconcentrate 12 is in this embodiment is contained in a dissolvingcapsule 15 designed to dissolve during mixing with the diluent 13. The diluent 13 may be acidic or basic depending on the chemistry of the concentrate, and thecapsule 15 formed of a material which dissolves in the diluent 13. The diluent 13 may be contained in a mixing well associated with acover member 14 of the SSM inFIG. 1A , or a mixing well, diluent chamber or mixing chamber of the instrument as illustrated inFIG. 2A andFIG. 2B . - In the embodiments of
FIG. 4B andFIG. 4C , the concentrate such as 12 is contained in aconcentrate dispenser 50 operatively coupled to an instrument for on-board provision of a reagent. Theconcentrate dispenser 50 includespackaging 52 within which one of a plurality of the concentrate capsules such as 12A to 12F are contained in a stacked cartridge. The stackedcartridge packaging 52 is designed to release each of the capsules such as 12A for mixing with a diluent on-board the instrument to provide a reagent at the required concentration. In the embodiment ofFIG. 4B theconcentrate dispenser 50 includes a reciprocatingrelease 54 operatively coupled to the stackedcartridge 52 to release a lowermost of the column of capsules such as 12A. - In the embodiment of
FIG. 4C , the packaging of theconcentrate dispenser 50 is in the form of acartridge reel 52 contained within the instrument. The cartridge reel contains a roll of the concentrate capsules such as 12A to 12F retained in a concentrate tape 56 dispensed from thecartridge reel 52. The concentrate tape 56 includes twostrips concentrate dispenser 50 ofFIG. 4C also includes aspool 60 within the instrument for removing thelower strip 58B of the concentrate tape 56 to release each of the concentrate capsules such as 12A from thedispenser 50. - In the embodiment of
FIG. 4D the instrument includes achamber 16 in the form of a disposable or reusable probe tip containing theconcentrate 12. Theprobe tip 16 is designed to fit to a probe such as 44 of the concentrate dispenser. The probe may for example be in the form of a FTP of an instrument used in treating or staining tissue samples. In this case theprobe tip 16 includes anozzle 46 designed to cooperate with a port of for example a cover member of a SSM. Thenozzle 46 includes atemporary seal 48 which is designed to rupture on discharge of theconcentrate 12 into the well for mixing with the diluent. Theconcentrate 12 may also be sealed in theprobe tip 16 with anotherseal 49 which is similarly ruptured under the application of pressure applied by theFTP 44. -
FIG. 5A ,FIG. 5B , andFIG. 5C illustrate various embodiments of adiluent chamber 17 operatively coupled to aconcentrate dispenser 19 for mixing on-board an instrument to produce a reagent. The diluent chamber is in the form of a mixingvial 17 with theconcentrate dispenser 19 attached as a single package. In the embodiment ofFIG. 5A thediluent vial 17 has a vacuum pressure applied to it for dispensing of theconcentrate 12 from theconcentrate dispenser 19. In the embodiment ofFIG. 5B theconcentrate dispenser 19 includes aplunger 21 for urging theconcentrate 12 from theconcentrate dispenser 19 into thediluent vial 17. Theconcentrate dispenser 19 may include a oneway valve 23 through which theconcentrate 12 is discharged into thediluent vial 17. The embodiment ofFIG. 5C includes a concentrate dispenser in the form of the packagedchamber 16 such as the embodiments ofFIG. 3A . Thechamber 16 is operatively coupled to the diluent chamber orvial 17 with its sealingmembrane 18 in contact with anentrance 25 to thediluent chamber 17. Theentrance 25 includes aspiked ridge 27 designed on the application of downward pressure to thechamber 16 to rupture thefrangible sealing membrane 18. Theconcentrate 12 is thus discharged into thediluent chamber 17 for mixing to produce the necessary reagent. - It will be readily understood by those skilled in the art that a concentrate includes liquids of varying concentrations and for example includes high-value reagents and bulk reagents. The FTP of the earlier embodiment may thus extend to dispensing a reagent to be diluted, a diluent, other reagents, or a combination of these liquids.
- The instrument according to various embodiments includes a controller configured to implement the preferred method of producing a reagent on-board the instrument. The method may be implemented via computer program code including instructions to control movement of a concentrate dispenser in, for example, dispensing a concentrate to a well of a cover member associated with a SSM as in
FIG. 1A , or a mixing well, diluent chamber or mixing chamber of the instrument as illustrated inFIGS. 2A and 2B . The controller may also control dilution and mixing of the concentrate with a diluent to provide the reagent at the required concentration. - Now that several preferred embodiments of the invention have been described it will be apparent to those skilled in the art that the method of producing a reagent on-board an instrument together with the concentrate dispenser and associated packaging has at least the following advantages:
- 1. The concentration of the reagent produced on-board the instrument can be varied, for example, to achieve the required staining option for tissue samples;
- 2. The ability to produce reagents on-board an instrument increase workflow flexibility with ease of use, for example by controlling the parameters of the dilution based on the supplied concentrate errors in staining are reduced;
- 3. On-board dilution lends itself to reagents in a liquid, solid and lyophilised form;
- 4. On-board dilution improves laboratory workflow without having to pre-dilute reagents from concentrates and also reduces operator time and potential errors.
- Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. All such variations and modifications are to be considered within the scope of the present invention the nature of which is to be determined from the foregoing description.
- Future patent applications may be filed in the United States of America on the basis of or claiming priority from the present application. It is to be understood that the following claims are provided by way of example only, and are not intended to limit the scope of what may be claimed in any such future application. Features may be added to or omitted from the claims at a later date so as to further define or re-define the invention or inventions.
Claims (15)
1. An instrument utilizing a reagent, said instrument providing on-board mixing of the reagent and including:
a probe for dispensing said reagent;
a concentrate chamber adapted to contain a concentrate; and
a diluent chamber adapted to contain a diluent;
wherein the probe includes an in-line mixing chamber adapted to receive the concentrate and the diluent to provide the reagent at the required concentration for dispensing by the probe.
2. The instrument as defined in claim 1 , wherein the concentrate chamber is located in the probe, upstream of the mixing chamber.
3. The instrument as defined in claim 1 , wherein the diluent chamber is located in the probe, upstream of the mixing chamber.
4. The instrument as defined in claim 1 , wherein the probe includes a bifurcation upstream of the mixing chamber providing fluid communication between each of the concentrate chamber and the and the diluent chamber.
5. The instrument as defined in claim 1 , wherein the diluent is pumped to the diluent chamber from a bulk supply on board the instrument.
6. The instrument as defined in claim 1 , wherein the concentrate chamber receives a predetermined volume of the concentrate from a sealed container stored on-board the instrument.
7. The instrument as defined in claim 6 , wherein the sealed container includes a sealing membrane arranged to contain the concentrate and an inert gas, and wherein the sealing membrane is adapted to be at least partly removed or pierced by the instrument.
8. The instrument as defined in claim 6 , comprising a plurality of the sealed containers on-board the instrument, each sealed container containing a predetermined volume of the concentrate.
9. The instrument as defined in claim 8 , wherein the plurality of sealed containers are arranged as multiple reservoirs in a blister pack.
10. The instrument according to claim 1 , including a concentrate dispenser comprising packaging adapted to contain a plurality of concentrate capsules on-board the instrument, and wherein the instrument is configured to control release each of the capsules from the concentrate dispenser for mixing with a diluent in the mixing chamber of the probe.
11. The instrument as defined in claim 10 wherein the packaging includes a stacked cartridge adapted to contain one or more columns of the plurality of concentrate capsules.
12. The instrument as defined in claim 11 wherein the concentrate dispenser also comprises a reciprocating release operatively coupled to the stacked cartridge to release, under control of the instrument, a lowermost of the column of capsules from the cartridge for mixing with the diluent on-board the instrument.
13. The instrument as defined in claim 10 , wherein the packaging includes a cartridge reel adapted to contain a roll of the concentrate capsules retained in a concentrate tape configured to be dispensed from the cartridge reel under control of the instrument.
14. The instrument as defined in claim 13 , wherein the concentrate tape includes two strips adhered to one another with the concentrate capsules sandwiched therebetween, and the concentrate dispenser includes a spool for removing one of the strips to release each of the concentrate capsules from the dispenser.
15. The instrument as defined in claim 1 , wherein the instrument is a laboratory instrument and the reagent is a high value reagent.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/066,094 US20210121884A1 (en) | 2012-12-21 | 2020-10-08 | Method of producing a reagent on-board an instrument |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201261745331P | 2012-12-21 | 2012-12-21 | |
PCT/AU2013/001499 WO2014094062A1 (en) | 2012-12-21 | 2013-12-20 | Method of producing a reagent on-board an instrument |
US201514653956A | 2015-06-19 | 2015-06-19 | |
US17/066,094 US20210121884A1 (en) | 2012-12-21 | 2020-10-08 | Method of producing a reagent on-board an instrument |
Related Parent Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/653,956 Continuation-In-Part US20150343445A1 (en) | 2012-12-21 | 2013-12-20 | Method of producing a reagent on-board an instrument |
PCT/AU2013/001499 Continuation-In-Part WO2014094062A1 (en) | 2012-12-21 | 2013-12-20 | Method of producing a reagent on-board an instrument |
Publications (1)
Publication Number | Publication Date |
---|---|
US20210121884A1 true US20210121884A1 (en) | 2021-04-29 |
Family
ID=75585196
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US17/066,094 Pending US20210121884A1 (en) | 2012-12-21 | 2020-10-08 | Method of producing a reagent on-board an instrument |
Country Status (1)
Country | Link |
---|---|
US (1) | US20210121884A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
USD999425S1 (en) * | 2019-11-25 | 2023-09-19 | Illumina, Inc. | Cartridge cover |
Citations (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5073505A (en) * | 1990-06-27 | 1991-12-17 | United Technologies Corporation | Eluant storage and preparation apparatus and method for using the same in a zero gravity environment |
US6211956B1 (en) * | 1998-10-15 | 2001-04-03 | Particle Sizing Systems, Inc. | Automatic dilution system for high-resolution particle size analysis |
US6360595B1 (en) * | 2001-03-16 | 2002-03-26 | Ethicon Endo-Surgery, Inc. | Liquid measuring device and method of using |
US20060013725A1 (en) * | 2002-06-11 | 2006-01-19 | Larsen Ulrik D | Disposable cartridge for characterizing particles suspended in a liquid |
US20100240022A1 (en) * | 2006-06-23 | 2010-09-23 | Mcneely Michael R | Reagent preparation and valving design for liquid testing |
US7832293B2 (en) * | 2003-06-17 | 2010-11-16 | Hemosystem | Method and device for drawing and mixing liquid samples |
US20100317093A1 (en) * | 2009-06-10 | 2010-12-16 | Cynvenio Biosystems, Inc. | Flexible pouch and cartridge with fluidic circuits |
US20130208558A1 (en) * | 2010-06-29 | 2013-08-15 | Biolyph Llc | Reagent preparation assembly |
US20140033809A1 (en) * | 2011-03-09 | 2014-02-06 | Pixcell Medical Technologies Ltd | Disposable cartridge for preparing a sample fluid containing cells for analysis |
US20160199834A1 (en) * | 2015-01-14 | 2016-07-14 | Pixcell Medical Technologies Ltd | Disposable cartridge for sample fluid analysis |
US20170058245A1 (en) * | 2015-08-27 | 2017-03-02 | Rushabh Instruments, Inc. | Immuno histo chemistry tissue processing system and cartridge therefor |
US20170274376A1 (en) * | 2016-03-28 | 2017-09-28 | Tangen Biosciences, Inc. | Apparatus and method for extracting pathogens from biological samples |
US20190277837A1 (en) * | 2017-06-02 | 2019-09-12 | Chengdu Polytech Biological Technology Co., Ltd. | Device and method for capillary chemiluminescence detection |
US20200179932A1 (en) * | 2012-02-13 | 2020-06-11 | Neumodx Molecular, Inc. | System and method for processing and detecting nucleic acids |
US20200316599A1 (en) * | 2017-10-23 | 2020-10-08 | Cytochip Inc. | Devices and Methods for Measuring Analytes and Target Particles |
US20200400534A1 (en) * | 2018-02-27 | 2020-12-24 | CytoChip, Inc. | Devices and methods for sample analysis with serial dilution |
US20220048031A1 (en) * | 2018-12-20 | 2022-02-17 | Alveo Technologies, Inc. | Methods and compositions for detection of amplification products |
US20220305493A1 (en) * | 2019-07-03 | 2022-09-29 | Robert Bosch Gmbh | Microfluidic Device for Processing and Aliquoting a Sample Liquid, Method and Controller for Operating a Microfluidic Device, and Microfluidic System for Carrying Out an Analysis of a Sample Liquid |
US20230010400A1 (en) * | 2019-12-04 | 2023-01-12 | Ian Basil Shine | Improved methods and devices for measuring cell numbers and/or cell properties |
-
2020
- 2020-10-08 US US17/066,094 patent/US20210121884A1/en active Pending
Patent Citations (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5073505A (en) * | 1990-06-27 | 1991-12-17 | United Technologies Corporation | Eluant storage and preparation apparatus and method for using the same in a zero gravity environment |
US6211956B1 (en) * | 1998-10-15 | 2001-04-03 | Particle Sizing Systems, Inc. | Automatic dilution system for high-resolution particle size analysis |
US6360595B1 (en) * | 2001-03-16 | 2002-03-26 | Ethicon Endo-Surgery, Inc. | Liquid measuring device and method of using |
US20060013725A1 (en) * | 2002-06-11 | 2006-01-19 | Larsen Ulrik D | Disposable cartridge for characterizing particles suspended in a liquid |
US7832293B2 (en) * | 2003-06-17 | 2010-11-16 | Hemosystem | Method and device for drawing and mixing liquid samples |
US20100240022A1 (en) * | 2006-06-23 | 2010-09-23 | Mcneely Michael R | Reagent preparation and valving design for liquid testing |
US20100317093A1 (en) * | 2009-06-10 | 2010-12-16 | Cynvenio Biosystems, Inc. | Flexible pouch and cartridge with fluidic circuits |
US20130208558A1 (en) * | 2010-06-29 | 2013-08-15 | Biolyph Llc | Reagent preparation assembly |
US20140033809A1 (en) * | 2011-03-09 | 2014-02-06 | Pixcell Medical Technologies Ltd | Disposable cartridge for preparing a sample fluid containing cells for analysis |
US20200179932A1 (en) * | 2012-02-13 | 2020-06-11 | Neumodx Molecular, Inc. | System and method for processing and detecting nucleic acids |
US20160199834A1 (en) * | 2015-01-14 | 2016-07-14 | Pixcell Medical Technologies Ltd | Disposable cartridge for sample fluid analysis |
US20170058245A1 (en) * | 2015-08-27 | 2017-03-02 | Rushabh Instruments, Inc. | Immuno histo chemistry tissue processing system and cartridge therefor |
US20170274376A1 (en) * | 2016-03-28 | 2017-09-28 | Tangen Biosciences, Inc. | Apparatus and method for extracting pathogens from biological samples |
US20190277837A1 (en) * | 2017-06-02 | 2019-09-12 | Chengdu Polytech Biological Technology Co., Ltd. | Device and method for capillary chemiluminescence detection |
US20200316599A1 (en) * | 2017-10-23 | 2020-10-08 | Cytochip Inc. | Devices and Methods for Measuring Analytes and Target Particles |
US20200400534A1 (en) * | 2018-02-27 | 2020-12-24 | CytoChip, Inc. | Devices and methods for sample analysis with serial dilution |
US20220048031A1 (en) * | 2018-12-20 | 2022-02-17 | Alveo Technologies, Inc. | Methods and compositions for detection of amplification products |
US20220305493A1 (en) * | 2019-07-03 | 2022-09-29 | Robert Bosch Gmbh | Microfluidic Device for Processing and Aliquoting a Sample Liquid, Method and Controller for Operating a Microfluidic Device, and Microfluidic System for Carrying Out an Analysis of a Sample Liquid |
US20230010400A1 (en) * | 2019-12-04 | 2023-01-12 | Ian Basil Shine | Improved methods and devices for measuring cell numbers and/or cell properties |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
USD999425S1 (en) * | 2019-11-25 | 2023-09-19 | Illumina, Inc. | Cartridge cover |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US9422067B2 (en) | Method of providing a chemical or biological material in quantised form and system therefor | |
US7789270B2 (en) | Chemical reaction cartridge and method using same | |
JP4546534B2 (en) | Comprehensive and automatic analyzer for DNA or protein in a disposable cartridge, method for manufacturing such cartridge, and operating method for DNA or protein analysis using such cartridge | |
CN1950520B (en) | Method and assembly for DNA isolation with dry reagents | |
RU2741391C2 (en) | Analytical device | |
CN103728461B (en) | Liquid relief pipe unit and the method for aspirating test liquid | |
JP4888394B2 (en) | Microreactor and liquid feeding method using the same | |
JP4593451B2 (en) | Microreactor system and liquid feeding method | |
US20070263049A1 (en) | Supply arrangement with supply reservoir element and microfluidic device | |
EP1873533A1 (en) | Testing chip for analysis of sample, and microanalysis system | |
US20210121884A1 (en) | Method of producing a reagent on-board an instrument | |
CN101878427A (en) | The micro-chamber plate, manufacturing method thereof | |
WO2020107641A1 (en) | Biological reaction device provided with microfluidic or nanofluidic structure | |
US20040048388A1 (en) | Microfluidic chip for biomolecule crystallization | |
JP2007083191A (en) | Microreacter | |
EP2934730B1 (en) | Method of producing a reagent on-board an instrument | |
CN106457248B (en) | Device, method and use for transferring liquids | |
JP2018141686A (en) | Liquid feeding method using specimen processing chip and liquid feeding device of specimen processing chip | |
US11426721B2 (en) | Bubble eliminating structure, bubble eliminating method, and agitating method using the same | |
JP2007083190A (en) | Microreacter | |
US20020137199A1 (en) | Micro storage, reaction and detection cells and method and apparatus for use thereof | |
JP3967331B2 (en) | Liquid mixing method, liquid mixing apparatus and microchip | |
AU2018250920B2 (en) | Device for processing a liquid sample | |
EP2409765A1 (en) | Microfluidic system and process for mixing liquid fluids | |
CN116569196A (en) | System and method for processing a sample |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: APPLICATION DISPATCHED FROM PREEXAM, NOT YET DOCKETED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: FINAL REJECTION COUNTED, NOT YET MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: FINAL REJECTION MAILED |