EP3532599A1 - Vorrichtung und verfahren zur behandlung biologische zellen enthaltender proben - Google Patents
Vorrichtung und verfahren zur behandlung biologische zellen enthaltender probenInfo
- Publication number
- EP3532599A1 EP3532599A1 EP17800378.6A EP17800378A EP3532599A1 EP 3532599 A1 EP3532599 A1 EP 3532599A1 EP 17800378 A EP17800378 A EP 17800378A EP 3532599 A1 EP3532599 A1 EP 3532599A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- sample
- sample carrier
- samples
- carrier
- receiving
- 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
- 238000000034 method Methods 0.000 title claims description 51
- 239000000969 carrier Substances 0.000 claims abstract description 39
- 210000004369 blood Anatomy 0.000 claims abstract description 7
- 239000008280 blood Substances 0.000 claims abstract description 7
- 238000003860 storage Methods 0.000 claims description 37
- 238000004458 analytical method Methods 0.000 claims description 33
- 238000009736 wetting Methods 0.000 claims description 31
- 238000005496 tempering Methods 0.000 claims description 14
- 239000000126 substance Substances 0.000 claims description 13
- 238000007599 discharging Methods 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 361
- 210000004027 cell Anatomy 0.000 description 100
- 239000000872 buffer Substances 0.000 description 54
- 239000000243 solution Substances 0.000 description 22
- 238000002360 preparation method Methods 0.000 description 20
- 238000010438 heat treatment Methods 0.000 description 15
- 238000002156 mixing Methods 0.000 description 15
- 238000001816 cooling Methods 0.000 description 14
- 238000001035 drying Methods 0.000 description 12
- 239000000203 mixture Substances 0.000 description 12
- 239000002671 adjuvant Substances 0.000 description 10
- 210000003850 cellular structure Anatomy 0.000 description 10
- 238000012545 processing Methods 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 239000003086 colorant Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 230000001681 protective effect Effects 0.000 description 6
- 210000000170 cell membrane Anatomy 0.000 description 5
- 230000007613 environmental effect Effects 0.000 description 5
- 230000001476 alcoholic effect Effects 0.000 description 4
- 239000012752 auxiliary agent Substances 0.000 description 4
- 239000000470 constituent Substances 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 230000009172 bursting Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000000383 hazardous chemical Substances 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012958 reprocessing Methods 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000001143 conditioned effect Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000009395 genetic defect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000000633 nuclear envelope Anatomy 0.000 description 1
- 239000000123 paper Substances 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/02—Means for pre-treatment of biological substances by mechanical forces; Stirring; Trituration; Comminuting
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/02—Filters
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/14—Incubators; Climatic chambers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/06—Means for pre-treatment of biological substances by chemical means or hydrolysis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/20—Heating; Cooling
Definitions
- the invention relates to an apparatus and a method for treating samples containing biological cells, in particular blood or cell samples.
- the importance of genetic and molecular biological analyzes is growing in the life sciences sector and in the medical environment. Particularly in the areas of prognosis, diagnosis and documentation of, for example, genetic defects or tumor diseases, methods for reprocessing cell samples with the aim of making cell components analytically accessible have become indispensable. Part of this analysis is based on the destruction of the cell membrane by dropping a pretreated cell sample on a sample carrier. The cell membrane breaks and leaking cell components - including chromosomes - are used for subsequent procedures, eg. B. dyeings, and analysis steps, eg. As an image analysis, accessible.
- the manual preparation of a cell sample for analysis of components of the cell sample usually comprises the following steps:
- Sample preparation The cell sample whose cells have cell components to be subjected to analysis is usually in the form of raw sample material. First, it is necessary to recover the cell sample from the raw sample material. Depending on the cell type, this step may require different procedures. However, it is generally in the enrichment and purification of the raw sample material by z. As centrifuging and optionally further steps, such as a cultivation of the cells to generate the cell sample to be analyzed. For example, it is necessary for the preparation of a chromosome preparation to stimulate cell division and to interrupt during nuclear division.
- sample processing The cells of the cell sample obtained in step (i) are processed to allow the cell membrane to be broken later.
- suitable methods z As the cell envelope, ie depending on the type of cell For example, the cell wall, the cell membrane or the nuclear membrane are removed. Depending on the cell constituents to be analyzed, it may be provided to fix the cell sample.
- suitable buffer z. As methanol and acetic acid, the nature of the cell membrane is changed so that it ruptures or bursts under the action of a force. A buffer or mixtures of several buffers can be used. The type of buffer used depends on the type of sample and the analysis to which the components of the processed cell sample are to be subjected.
- optional Density Adjustment In some cases it may be desirable to adjust the cell density in the cell sample.
- Sample carrier preparation The sample carrier is usually wetted with a buffer solution in order to positively influence the distribution of the exiting cell constituents.
- the sample carrier is usually one
- drying and immobilization It is in some cases expedient to dry the cell sample on the sample carrier. The buffer evaporates. In this way, the cell sample, including the released cell components, can be immobilized for subsequent analysis.
- step (vi) or step (vii) may be subjected to analysis.
- further staining steps or reactive solutions may be used. be used before analysis of the cell sample z. B. by a microscopic examination is possible.
- the result and quality of analysis of the manually processed cell sample are influenced by factors related to the execution of the cell sample preparation steps.
- the preparation of the cell sample is such a sensitive process that, in addition to the skills of the performing laboratory technician, the environmental conditions affecting the cell samples and liquids consisting of one or more components affect not only the processing itself but also the analysis. Not least because of the high volume of samples in the laboratory, it is often not possible to wait for the process environment to stabilize before starting the sample preparation. The multiple analysis of a sample in the same laboratory can therefore already lead to qualitatively different results.
- the object of the invention is to eliminate the disadvantages of the prior art.
- a device for treating samples containing biological cells is to be specified, which offers high stability, high reliability and good reproducibility of the processed samples obtained using the device. It is also intended to provide a method by which samples containing biological cells can be treated.
- a device for the automated treatment of cell-containing samples, in particular of blood or other cell samples is provided.
- the device has at least the following components:
- a sample receiving device for receiving one or more samples
- a tool-receiving device for receiving one or more tools; a sample carrier receiving device for receiving one or more sample carriers; a discharge device for dropping one or more samples from a discharge position; and
- a catcher for catching one or more dropped samples in a collection position to obtain one or more prepared samples.
- the distance between the discharge position and the collection position which is also referred to below as the discharge height, is adjustable to a predetermined height.
- the device furthermore has at least one tempering device for tempering at least one of the sample carriers. It can be provided that one or more sample carriers by means of a temperature control, such as a heating and / or cooling element, are directly controlled. Alternatively or additionally, it can be provided that the one or more sample carriers are arranged in a tempered region of the device. Preferably, the sample carrier is cooled until the sample is applied to it. It can then be provided that the sample carrier is heated.
- the tempering device may have an adjusting unit for setting a predetermined angle of inclination of the sample carrier.
- a single temperature control device can be provided, which allows temperature control, ie, cooling and heating of the sample carrier in a single position.
- This is the common heating and cooling position.
- It may be a heating and cooling element, such as a heating and cooling block, act.
- two tempering devices can be provided, with a first tempering device in a first position, for example for cooling the sample carrier before and during ejection of the sample, and a second tempering device in a second position, for example for heating the sample carrier after ejecting the sample Sample is arranged.
- the first Per ists shark may be a cooling element, such as a cooling block
- the second tempering a heating element such as a heating block
- the device according to the invention can be an automated device.
- the discharge device can have a pipetting system with a pipetting head, which is movable at least along one axis, preferably two axes and particularly preferably in the x, y, z direction.
- the discharge unit may have robotics.
- the robotics can be controlled by means of a control device.
- the ejection device may be a pipetting robot.
- the control device may be a data processing unit.
- the data processing unit is expediently arranged outside the housing, but may also be integrated into the housing or the device according to the invention. It can communicate with the ejection device via a data connection, for example a cable.
- the pipetting head may carry dosing agents, such as pipettes, capillaries or syringes.
- the dosing agents allow the inclusion of a predetermined amount of the sample from the sample containers.
- the discharge device can have at least one pipetting channel for taking a predefined amount of the sample from a sample container and for guiding the sample into the discharge position.
- the pipetting system can also be used to mix the buffer from auxiliaries.
- a separate mixing device and guide means for guiding one or more aids to the mixing device are then not required.
- the pipetting system can be used for wetting the sample carrier with one or more auxiliary means, for example a buffer.
- a separate wetting device and guide means for guiding one or more auxiliary Medium to the wetting device are then not required.
- One or more aids, for example a buffer can be dropped onto the sample carrier by means of the pipetting system. Also the setting of the - explained below - sample concentration is possible via the pipetting system.
- the pipette may have a replaceable tip or a removable needle.
- the needle may be a hollow needle.
- the needles can be washed.
- the device according to the invention can have a holding device for the pipette tips and / or needles, which is referred to below as a pipette tip holding device.
- the pipette tip holding device can be arranged in the device, preferably in its working area. It can be provided that samples, sample carriers and, if the use of one or more aids is provided, the aid or aids to a defined temperature below the ambient temperature, d. H. the room temperature, cooled before the sample is dropped onto the sample carrier.
- an adjuvant such as a buffer
- the invention is directed to the treatment of biological samples containing cells.
- treatment may include the preparation of the samples for subsequent analysis or the preparation and analysis of the samples, which in both cases may be, for example, a genetic, a molecular biological examination or both.
- the device according to the invention can have, as a further component, an analysis device for analyzing the discarded sample located on the sample carrier.
- the sample is not only processed, but also subjected to analysis within the device.
- the analysis device can be an imaging device, for example a microscope with a camera, as well as an image-evaluating unit, for example a staff Computer with image processing software, acting.
- the imaging device enables the acquisition of a high-quality image of the sample containing biological cells, the evaluation of which can be done manually or image-analytically by means of image processing software.
- the analysis can be directly connected to the sample preparation.
- the cells that may be included in the sample are, for example, eukaryotic cells.
- the invention makes it possible to make cell components of the cells contained in the sample accessible, for example visible, for subsequent analysis.
- the sample may be, for example, a blood sample.
- the sample is also referred to below as a cell sample.
- the distance between the dropping position and the catching position can be set to a value between 0.1 and 800 mm.
- the distance between the dropping position and the catching position is also referred to below as the discharge height.
- the inclination angle of the sample carrier to a value between 0 and 90 °, relative to a horizontal plane, is adjustable. Inclination angles of 15 to 75 ° are preferred.
- the collecting device has a holder for receiving one of the sample carriers. The collecting device may be guided so that the sample carrier located in the holder reaches the collecting position.
- the collecting device may have an adjusting unit for setting a predetermined angle of inclination of the sample carrier.
- the sample is dropped in the form of a drop.
- the flow of the drop of the sample is dependent on one or more of the factors sample type, analysis target and impact velocity, although the list is not exhaustive.
- the collecting device may be located at a fixed location or be movable along at least one axis, preferably two axes and more preferably in the x, y, z direction. At least one of the axes should be a horizontal axis.
- the device according to the invention can have a handling unit which can remove one or more sample carriers from the sample carrier receiving device and transfer them to the collecting device.
- the collecting device can have a handling unit which can be used to remove a Benffys from the sample holder receiving device is capable of and leads the sample carrier in the collection position.
- the handling unit can bring about a temperature control of the samples.
- the inclination of the sample carrier and / or the discharge height of the sample drop can be adjusted by the handling unit.
- the handling unit can be used to dip the sample carrier in a buffer or to position the sample carrier so that buffer can be applied by means of the pipetting system.
- one or more protective elements may be arranged to effect a splash guard.
- the discharge path which travels the sample from the discharge position to the collection position, wholly or partially between such protective elements.
- the protective element may be, for example, a pipe through the interior of which the discharge path runs.
- partitions may also be provided as protective elements between the sample carriers, which prevent contamination of the adjacent regions with sample spatters.
- These protective elements can be designed so that they can be washed and sterilized or used as disposable products, so-called disposable products. For example, the protective elements of Kunststoffsoff, paper, cardboard, etc. exist.
- the slides can be slides, such as slides called glass slides.
- the sample carrier is optional during collection of the sample in a refrigerated state. It can either be actively cooled or positioned on a cold block or covered with cold buffer. For coating the sample carrier with cold buffer, the sample carrier can be immersed, for example, in the buffer.
- the device according to the invention can have a wetting device for wetting the sample carrier with one or more auxiliary means.
- one or more auxiliary means are tempered by means of a tempering device, for example a cooling element. That is an immediate tempering.
- the aid or aids are arranged in a tempered area.
- the temperature-controlled area in which the sample carriers are arranged and the temperature-controlled area in which the aid or aids are arranged can be one and the same temperature-controlled area.
- the temperature-controlled area may be the tempered storage area described below, which is formed together with a working area in a housing.
- the aid or aids can be arranged instead of in the tempered storage area in a tempered partial work area of the work area. Wetting may not be required if coated sample carriers are used.
- the coated sample carrier may carry a coating that allows temporary or permanent immobilization of components of the ruptured sample on the sample carrier.
- the sample carrier may be coated with antibodies.
- proteins from the ruptured sample can be immobilized on the sample carrier.
- the sample carrier may be coated with proteins. In this way, for example, antibodies from the ruptured sample can be immobilized on the sample carrier.
- the sample receiver may be a rack.
- the sample receiver may receive one or more samples. It can be provided that the samples are introduced into the sample receiving device via the feed, which are formed in the housing. As an alternative, it can be provided that the sample receiving device is introduced as such via the feed, which is formed in the housing. In this case, the sample or samples are introduced into the sample receiving device prior to inserting the sample receiving device into the housing. Conveniently, the samples are in sample containers when introduced into the sample receiver. In this case, the sample container can either be inserted in the already open state or automatically opened by an opening device of the device according to the invention.
- the aid receiving device serves to receive one or more aids.
- the device according to the invention can have a plurality of auxiliary device receiving devices.
- An adjuvant is a buffer or substance needed to make a buffer.
- Such substances are, for example, an alcoholic solution such as a methanol solution and an acid solution such as an acetic acid solution.
- the aids are in aid containers when they are inserted into the aid receptacle.
- the aid containers can either be inserted in the already opened state or opened automatically by an opening device of the device according to the invention.
- the device according to the invention has as a further component a mixing device for producing a mixture of adjuvants.
- the mixing device may be arranged in the housing. However, the mixing device may also be part of a wetting device or a pipetting channel.
- the aid receiving device can be used to separately receive all substances that are required for the production of the buffer. These can be two or more substances.
- the buffer is mixed with two substances, an alcohol and an acid.
- the substances required for the preparation of the buffer can be mixed in the mixing device to form a mixture. This mixture can then be used as a buffer.
- any, but defined amount of the auxiliary can be performed to the mixing device.
- the quantities of the aids can be specified by the user.
- a mixing device is not required when using prefabricated buffers.
- a wetting device serves for wetting the sample carrier with one or more auxiliary means.
- the wetting device serves for wetting the sample carrier with a buffer.
- the wetting device can have one or more nozzles with which one or more aids, for example a buffer, are applied to the sample carrier.
- the wetting device may comprise a reservoir in which one or more aids, for example a buffer, are located and into which the sample carrier can be immersed.
- the wetting of the sample carrier serves for its pretreatment.
- One or more adjuvants may be mixed or sequentially applied to a sample carrier.
- the tool is a buffer, in another embodiment a combination of various tools, which may be different buffers.
- one or more guiding means can be provided.
- a guide means are a pump, a pump system, one or more washable needles, one or more pipettes.
- a pipette may have a replaceable tip.
- the sample carrier receiving device may be a rack.
- the sample holder receiving device can record the unused sample carriers, ie the sample carriers to which a sample is to be dropped, and the sample carriers used, ie the sample carriers onto which a sample has already been dropped.
- the sample carrier receiving device can be tempered, preferably cooled.
- the device according to the invention may have a device for determining the cell density in the sample. This device is referred to below as Zellêt- measuring device.
- the determination of the cell density in the sample may be provided to set a predetermined density of cells in the sample before it is discarded. If the cell density is above a predetermined value, for example, it may be provided to add a buffer or a buffer mixture to the sample in order to set a defined cell density.
- One or more guiding means may be provided for guiding the aid (s) from the aid receiving means to the sample container in which the sample is located. These guiding means are also referred to as tracking means.
- the cell density in the sample reflects the concentration of cells in the sample, which is why it is also referred to as cell density concentration or sample concentration.
- the measurement of the cell density can be used to set further process parameters. These include one or more of the following parameters: angle, ambient conditions, temperature of the sample carrier, temperature of the environment and the discharge height.
- the device according to the invention has a storage area in which at least the sample carrier receiving device is arranged.
- the storage area may have the ambient temperature or tempered, preferably cooled.
- the storage area is then a tempered area.
- the device according to the invention has a temperature-controlled, preferably cooled storage area, it is also possible for the auxiliary device receiving device to be arranged next to the sample carrier receiving device in the temperature-controlled area.
- the device according to the invention has a working area in which at least the discharge device and the collecting device are arranged.
- the working area can be tempered, for example cooled or heated.
- the working area can be regulated to a specific humidity.
- the sample receiving device is arranged in the work area.
- the aid receiving device is preferably arranged in the working area.
- a tempered partial work area can be formed in the work area.
- the tempered partial work area is preferably a cooled area.
- one or more auxiliary-receiving devices may be arranged.
- the aids received by these auxiliary receiving devices can be tempered, preferably cooled.
- the sample receiving device may be arranged.
- the storage area and the work area may be formed in the housing. Conveniently, the work area adjoins the storage area.
- the device according to the invention comprises a housing in which one or more components of the device are arranged. In the housing, at least one space can be formed in which physical and / or chemical conditions that are different from those of the environment can be set. This particularly applies to one or more of the following Conditions: temperature, pressure and composition of the gas phase. This gas phase is also referred to below as working gas.
- the room may be the storage area, the partial work area, the entire work area or the entire interior of the enclosure. It can be provided that the temperature and / or the humidity, preferably both, in the region of the housing differ from those of the environment. Preferably, the temperature is below the ambient temperature.
- the ambient temperature is usually the room temperature. It can be provided that the temperature is at least in a tempered range in the housing below 0 ° C, for example at -20 ° C. It can be provided that the air humidity is adjusted depending on the sample type, the pretreatment and the ambient conditions. It can be provided that the working gas is air.
- the device according to the invention may have a device for the treatment of the working gas, which generates a purified homogeneous gas stream.
- the means for treating the working gas may be a filter.
- the device according to the invention may further comprise means for guiding the working gas to the means for treating the working gas and away from the means for treating the working gas.
- the means for guiding the working gas may be a fan.
- the physical and / or chemical conditions in the device or in a region of the device are also referred to below as the climate.
- at least one conditioned sterile room is formed in the housing. It can be the workspace, the storage area or both.
- a laminar flow of a working gas may be formed in the housing.
- the following components of the device according to the invention can be arranged in the housing: at least one auxiliary device receiving device, the sample carrier receiving device, the discharge device and the collecting device.
- the wetting device for wetting the sample carrier with one or more auxiliary means, if provided, may also be arranged in the housing.
- the analysis device can also be arranged in the housing.
- the device according to the invention may have a feed for introducing one or more samples into the housing.
- the sample receiving device may be disposed in the housing. It may be provided that the subcontractor Work area is formed in the housing so that the samples can be introduced via the feed in the sub-work area.
- the housing By means of the housing, in particular for clinical applications advantageous enclosure of the components of the system, which come into contact with the sample can be achieved. Having an internal, homogenous, purified airflow into the housing can provide the necessary sterility of the sample and process environment. It can be provided to connect the device according to the invention to a trigger system in which filtered exhaust air can be discharged from the interior of the housing. This may be appropriate or required by law, especially when using potentially hazardous substances or combinations that form hazardous substances.
- the device according to the invention has at least one identification device (eg barcode reader) with which identification means (eg barcodes) which are located on the sample container, sample carrier and / or reagent container can be identified .
- the identification device eg barcode reader
- the identification device can also be used to identify consumables, for example pipette tips or their storage containers, thus making it easier to allocate samples and sample carriers, as well as allowing process control, which is also referred to as tracking.
- the identification means (eg barcode) may be, for example, optical codes, in particular a barcode, or RFID tags or other identification means.
- a method of automated treatment of biological cell-containing samples particularly blood or other cell samples. In the method, a temperature-controlled sample carrier is wetted with one or more tempered auxiliaries and a sample of a predetermined height is dropped on the wetted sample carrier.
- the temperature-controlled sample carrier and the tempered or the auxiliaries are heated to a temperature below the ambient temperature, more preferably below 0 ° C and particularly preferably -20 ° C.
- the The sample is heated to a temperature below room temperature before being dropped onto the temperature-controlled sample carrier.
- the temperature-controlled sample carrier is tilted by a predetermined inclination angle to a horizontal plane before the sample is dropped onto the temperature-controlled sample carrier. It can also be provided that the sample is subjected to an analysis after being dropped onto the temperature-controlled sample carrier. It can be provided that one or more catchment areas are formed on a sample carrier. In this case, one or more drops of the same sample or from different samples can be dropped on each collection area. The sample carrier must be aligned in such a way that the collecting area to which one or more drops are to be applied is in the collecting position. Multiple collecting areas on a sample carrier allow better utilization of the surface of the sample holder. Advantageously, several drops are dropped onto different catchment areas of the sample carrier, it being important to ensure compliance with the discharge height, based on the inclination of the sample carrier.
- a sample carrier can also have a collecting position which, although treated with all process liquids, is not provided with a sample. This collection area can then serve as a negative control. In this way, possibly existing contamination of the aids and / or the sample carrier can be detected.
- the sample carrier can be adjusted to a predetermined specific angle of inclination.
- the angle of inclination can be adjusted between 0 and 90 ° to the horizontal plane. In this case, the surface of the sample carrier on which the sample is to be dropped is in a horizontal plane when the angle is 0 °. It lies in a vertical plane when the angle is 90 °. At an angle of 90 °, the sample holder is thus vertical.
- the angle of inclination may be specified by the user depending on the planned analysis of the sample and the type of sample.
- process parameters that is, for example, one or more of the factors environmental conditions, quantities of auxiliaries, quantities of media, discharge heights, discharge speeds and angles of inclination, can also be defined by a predetermined protocol.
- a protocol can only be used by non-specialist and competent administrators of normal users and can not be manipulated. This increases process reliability and prevents errors due to incorrect process parameters.
- the protocol preferably defines at least discharge heights, discharge speeds and inclination angles.
- a defined amount of the sample is taken from the sample container containing a sample by means of the discharge device.
- the sample may be a prepared sample.
- the cell density of the sample is determined and, if the cell density exceeds a predetermined value, adjusted using one or more auxiliaries, for example a buffer. It may further be provided that the sample has been identified by means of an identification means (eg barcode), which the sample container has, prior to the removal of the sample.
- the discharge device is a pipetting unit, for example a pipetting robot, it can be provided that the predetermined amount of sample is removed from the sample container by means of a pipette tip or needle. The pipette tip or needle is then aligned so that the drop is dropped from the dropping position.
- the required discharge height can be shortened by increasing the discharge rate of the cell sample at the moment of leaving the pipette tip or needle.
- the discharge height can be set to a value in the range of 0.1 to 800 mm.
- a break of defined length can be provided.
- the cell contents of the burst cell can run on the inclined sample carrier.
- This process can be assisted or positively influenced by a homogeneous gas flow.
- it may be provided to evaporate liquids that are on the sample carrier, in particular aids such as the buffer.
- the temperature at the sample carrier surface for example by means of a drying device, can be significantly increased, as a result of which these liquids evaporate.
- a drying at ambient temperature can be achieved. In both cases, drying can be accelerated through a ventilated environment.
- liquid adjuncts e.g. B to apply a dye to the sample carrier to stain or otherwise react some cell components.
- either one or more further dedicated pipette channels may be used as the guide means, or an existing pipette channel may be rinsed with an adjuvant and used to apply the liquid adjuvant. If necessary, a second drying can be carried out.
- the specimens located on the sample carrier and, if provided, immobilized samples can be subjected to an analysis, for example by means of a microscope.
- Use pipette tips may be discarded, used pipette needles, for example, by a cleaning method, such as washing with a washing solution, by plasma, heating or combinations thereof, cleaned and, if appropriate, wishes to be sterilized before further samples are processed in the same way.
- the method according to the invention can be carried out in particular by means of the device according to the invention. Further features of the method according to the invention will become apparent from the above description of the device according to the invention.
- the inventors have found that some environmental factors are critical to successful and traceable sample preparation. In particular, the inventors have determined the following environmental factors that have a significant influence on the results of the sample preparation:
- a low temperature of the cell sample, buffer and sample carrier promotes the bursting of the processed cells of the cell sample.
- the cells become more sensitive to shear and compressive forces acting on the cells as they impact the slide. This leads to an increased number of lysed cells with the same sample volume, which leads to an increased exposure of components of the cells of the cell sample.
- the ingredients are the material of interest of a subsequent analysis.
- a buffer usually contains several components, eg. As an acid component and an alcoholic component. These components often tend to segregate, so that before the preparation for good mixing of the buffer o-, if more buffers are used, the buffer must be respected. Uniform mixing of the components is easier to guarantee mechanically than with manual reprocessing. Further important factors for the success of the cell lysis and the spread of the cell components are the adherence to a defined dripping height and the adherence to a defined angle of inclination of the sample carrier. Another important factor is ensuring a uniform cell count of the cell sample. The cell count must be high enough to provide enough evaluable material. However, too high a cell number affects the readability z.
- prepared cell samples are obtained which are of high quality, comparable and validatable. They are particularly suitable for genetic and / or molecular biological analyzes.
- the invention makes it possible to increase the sample throughput compared with manual processing. This is in contrast to the increasing number of samples to be tested, with only slowly growing laboratory capacities and thus increasing waiting times for the provision of analysis material.
- uniform standards for sample preparation of genetic and / or molecular biological analyzes which correspond to scientific and clinical requirements, can be achieved for the first time on the basis of defined, consistent and monitored parameters.
- the parameters include in particular one or more, preferably all of the following parameters: a controlled temperature range in the storage area, a controlled temperature range in the working area rich, a constant air humidity in the work area, a precisely determined discharge height, a precisely measured discharge quantity, a precise adherence to the inclination angle of the sample carrier, a monitoring and compliance with the course and drying times and environmental conditions.
- Another optional parameter is a precisely rated discharge speed.
- the invention may be a splitting of cells and the fixation of the cell components flowing out of the cells, such. B. chromosomes, while offering a high stability and reproducibility, what the quality and significance of a subsequent, z. B. optical, evaluation increases.
- Fig. 1 is a plan view of a first embodiment of an inventive
- FIG. 2 shows a schematic illustration of the first embodiment of the device according to the invention without housing
- FIG. 3 shows a first flowchart for illustrating a first method sequence using the first embodiment of the device according to the invention.
- FIG. 4 shows a second flowchart for illustrating a second method sequence using the second embodiment of the device according to the invention.
- the first embodiment of the device according to the invention shown in FIGS. 1 and 2 has a housing 1.
- a cooled storage area 2 and a work area 3 are formed as a storage area.
- the storage area 2 and the work area 3 are chambers within the housing 1, which are connected to each other via an opening or a lock.
- a climate chamber 4 is disposed in the housing, in which there are means for processing and management of a working gas.
- the working gas is the gaseous medium in the housing, for example air.
- the means for processing the working gas the working gas can be treated, for example filtered.
- the means for treating the working gas may be, for example, a filter.
- the working gas from the cooled storage area 2, the work area 3 or both can be performed in the climatic chamber 4 to the means for processing the working gas and processed there.
- the treated working gas can then be returned to the refrigerated storage area 2, the work area 3 or both.
- a data processing device 5 for example a computer, is arranged, which serves to control the components of the device according to the invention arranged in the housing.
- a sample feeder 7 in the housing is a sample feeder 7 (see Fig. 3), which allows the introduction of samples in the work area 3.
- the cooled storage area 2 has a temperature of -20 ° C to room temperature.
- the working area 3 has a temperature of -20 C ° to room temperature.
- a sample carrier receiving device 21 is arranged, which allows the inclusion of a plurality of sample carriers 6.
- the sample carrier receiving device 21 may be a rack.
- a first auxiliary receiving device 22 for receiving a first auxiliary means and a second auxiliary receiving device 23 for receiving a second auxiliary means are provided.
- the first aid receiving device 22 and the second auxiliary receiving device 23 may be containers.
- the first aid may be a first substance needed to make a buffer.
- the first substance may be an alcohol.
- the second aid may be a second substance needed to make a buffer.
- the second substance may be an acid.
- a third auxiliary receiving device 24 (see Fig. 3) arranged to receive a third auxiliary means.
- the third aid Receiving device 24 may be a container.
- the third adjuvant may be, for example, a colourant or a reactive solution. It may be arranged in the refrigerated storage area 2 more auxiliary receiving devices. It should be expressly pointed out that it is not absolutely necessary to produce a buffer in the device according to the invention, for example from the first aid and the second aid. Rather, the buffer can already be introduced in ready-to-use form in the cooled storage area 2 in a tool-receiving device.
- the samples are conveniently in sample containers, for example in test tubes.
- the aids are expediently in aid containers.
- the sample containers, sample carriers 6 and / or the auxiliary containers may be labeled with identification means (eg barcode) which allow unambiguous identification of each individual sample, of each individual sample carrier and, if provided, of each individual aid. This permits the assignment and tracking of samples, sample carriers 6 and aids.
- the identification means eg barcode
- the identification means may be, for example, optical codes, in particular a barcode, or RFID tags or other identification means.
- the identification means (eg barcodes) can be applied by known methods to the sample containers, the sample carriers 6 and the auxiliary containers.
- the identification means make it possible to document the individual method steps comprehensively and together with the process parameters, for example in protocols.
- a wetting device 31 is provided for wetting sample carriers 6.
- the wetting device 31 has a mixing device (not shown) for producing an adjuvant mixture.
- the wetting device 31 also has nozzles 32, via which the auxiliary mixture can be applied to the sample carrier 6.
- the nozzles 32 may be one or more needles, in particular one or more washable needles, or one or more pipettes, in particular one or more pipettes with replaceable tips.
- a pump are provided for example hoses, and a pump are provided. Alternatively to the pump can Pump system be provided.
- the buffer can already be introduced into ready-to-use form in the cooled storage area 2 in a tool receiving device.
- no mixing device is required.
- the buffer from the auxiliary agent receiving device, in which the ready-to-use buffer is located can be guided via a guide means to a wetting device which has no mixing device.
- auxiliary agent or a mixture of adjuvants in a defined mixing ratio can be applied to a sample carrier 6.
- a collecting unit 40 for example a shell, is arranged, which receives the part of the aids or of the auxiliary agent mixture which does not adhere to the sample carrier 6.
- a collecting device 34 For removing the sample carrier 6 from the sample carrier receiving device 21, a collecting device 34 is provided.
- the collecting device 34 has a holding unit 35 for the sample carrier 6 and an adjusting unit 36 for inclining the sample holder 6 located in the holding unit 35.
- the holding unit 35 is movable in the x, y, z direction.
- the holding unit 35 and the adjusting unit 36 may be attached via an arm 37 to a rotor 38 which runs on a rail 39.
- the rotor 38 can be moved on the rail 39, for example by means of a servomotor, in a horizontal direction (x-coordinate).
- the rail 39 is supported in the housing 1 (not shown) so that the rail 39 is movable in the vertical direction (z coordinate) and a second horizontal direction (y coordinate).
- a pipetting unit 41 serving as a discharge device is arranged in the working area 3.
- the pipetting unit 41 has a pipetting head 42, which is movable in the x, y, z direction.
- the pipetting head 42 can be attached via an arm 43 to a rotor 44, which runs on a rail 45.
- the rotor 44 can be moved on the rail 45, for example by means of a servomotor, in a horizontal direction (x-coordinate).
- the rail 45 is mounted in the housing 1 (not shown) so that the rail 45 in the vertical direction (z-coordinate) and a second horizontal direction (y-coordinate) is movable.
- the pipetting head may hold one or more pipettes 46.
- the pipettes may have pipette tips or needles.
- the pipette tips of the pipettes 46 are changeable.
- the pipette needles of the pipettes 46 are changeable.
- the pipette needles can be washable.
- a pipette tip holding device 47 is further provided, which can receive one or more pipette tips or needles.
- a sample receiving device 48 may be arranged, which can take one or more samples.
- the sample receiving device 48 may be tempered, preferably cooled.
- the sample receiving device 48 may include a cell density measuring unit 49, with which a determination of the cell density of the samples prior to discarding is possible.
- a collecting container 49 may be arranged for used pipette tips or needles.
- the collection container 49 may include a dropper mechanism for separating the pipette tips or needles from a pipette.
- One or more identification devices 50 may be arranged in the work area 3, which allow identification of identification means (eg bar code).
- a drying device 51 for example a heating device, can be arranged in the working area 3, which enables drying and thus immobilization of the sample dropped on the sample carrier.
- an analysis device 52 for example a microscope, can be arranged in the working area 3, which allows an analysis of the sample dropped on the sample carrier and, if provided, dried sample.
- FIG. 3 shows the cooled storage area 2 and the work area 3. It should be noted that the refrigerated storage area 2 is shown twice in FIG. 3 because it holds both the empty sample carriers 6 'and the storage space
- the "process flow” arrow illustrates the procedure used to measure the sample and the sample carriers 6 "carrying the discarded samples. run.
- the individual process steps are identified in the flowchart by capital letters which are located in a circle.
- the method sequence is explained on the basis of a sample and with reference to a sample carrier 6. However, it is also possible to simultaneously handle a plurality of samples simultaneously and / or several sample carriers 6, even if this is not expressly stated. In addition, several samples can be treated successively by means of the device according to the invention. For this, the treatment of a previous sample does not have to be completed. The time interval between two samples may depend on the waiting time and / or drying time needed to treat a sample.
- sample carriers 6 there are empty sample carriers 6 'in the sample carrier receiving device 21, the aids required for producing the buffer, in particular the first and second aids, in the first or second auxiliary device receiving means 22, 23. Furthermore, there are pipette tips or needles in the pipette tip holder 47.
- the aids may be the following:
- Second aid Acid solution as a second buffer component
- the empty sample carriers 6 are uniquely identified by identification means (eg barcode).
- the containers in which the aids are located are clearly identified by means of identification (eg barcode).
- the cooled storage area 2 is cooled to a temperature of -20 ° C to room temperature. In the working area a defined climate is set, namely -20 ° C to room temperature. It can be provided that all settings of the device as well as the process sequences, in particular the process parameters and sample information, are logged.
- the catcher 34 is shown with a double arrow to illustrate the movement of the catcher 34 in a horizontal direction.
- the double arrows d and e illustrate the movement of the catcher 34 in a vertical direction and another horizontal direction.
- the collecting device 34 is thus movable in the x, y and z directions.
- the movement of the pipetting unit 41 in a horizontal direction is indicated by double arrow a.
- the movement of the pipette 41 in the vertical direction by the double arrows b.
- the pipetting head 42 is movable in the x, y and z directions.
- the method according to the invention starts when the sample carrier receiving device 21 is filled with clean sample carriers and one or more samples, which are located in sample containers, are introduced into the device according to the invention via the sample feed 7 in the housing 1 into the sample receiving device 48 in the work area 3 become.
- the samples may have previously been pretreated, for example centrifuged. They may also have been previously cooled to a temperature below room temperature.
- the sample or samples are contained in sample containers that are uniquely identified by means of identification (eg barcode).
- the sample containers can be opened or closed. It can be provided that the sample containers are located in a rack which is introduced into the working area 3 via the sample feed 7. It can be provided that several racks are introduced simultaneously into the work area 3.
- the samples are further cooled after introduction into the sample receiving device 48, at least as long as they are in the sample containers.
- Step A The sample or samples introduced into the sample receiver 48 are identified by an identifier 50 (eg, bar code reader). This should allow a later assignment of the sample to a sample carrier 6.
- Step B This step is optional.
- the cell density of the sample is determined by the cell density meter. It can be provided that the cell density of the sample is adjusted when the measured cell density falls below or exceeds a predetermined value.
- the cell density can be determined, for example, with an optical sensor and adjusted by means of the pipetting system and the buffer It can be provided that the sample is dyed with a colorant and / or with a reactive solution The colorant and the reactive solution may be stored as an aid in auxiliary receptacles in the refrigerated storage area 2.
- Step C The pipetting unit 41 is guided to the pipette tip holding device 47 and receives there one or more pipette tips or needles.
- the pipette tips or needles are used to collect samples later.
- Step D The pipetting unit 41 is guided to the sample receiver 48.
- a pipette 46 of the pipetting unit 41 a defined amount of a sample is taken from a sample container. The sample enters the pipette tip or needle of the pipette 46.
- Step E The catcher is moved into the refrigerated storage area 2.
- a sample carrier 6 ' is removed from the sample carrier receiving device 21, wherein the sample carrier 6' enters the holding unit 35.
- the sample carrier 6 ' is then guided by means of the collecting device 34 to the wetting device 31. He passes from the cooled storage area 2 in the work area. 3
- Step F The sample carrier 6 'is guided by means of the collecting device to a second identification device 50 (eg barcode reader) and identified there. This should allow a later assignment of the sample to the sample carrier 6.
- a second identification device 50 eg barcode reader
- Step G The first and second aids are mixed to form an adjuvant mixture.
- the adjuvant mixture is the buffer.
- defined amounts of the first and second auxiliary means from the first and second auxiliary receiving means 22, 23 are guided via the guide means 33 to the wetting device 31 and mixed there to form a buffer.
- the defined amounts of the first and second aids may correspond to a fixed predetermined mixing ratio.
- step B the cell density of the sample can be taken into account when specifying the defined amount.
- step G is only required if no prefabricated buffer is used. In this case, the buffer from a tool-receiving device in which it is stored refrigerated, guided over the guide means 33 to the wetting device 31, where it can be readily used.
- the auxiliary mixture can be prepared and the samples can be wetted with drops of the auxiliary mixture. This eliminates the wetting device 31, a mixing device or both.
- Step H The buffer is applied to the sample carrier 6 'via the nozzles 32.
- the sample carrier 6 ' is rinsed with the buffer and in particular wets the surface of the sample carrier 6' with the buffer.
- Several sample carriers 6 'can be rinsed simultaneously.
- the sample carriers 6 'can also be rinsed one after the other.
- Step I According to the specifications of the user of the device according to the invention and optionally taking into account the results of the cell density measurement, the following parameters are now set: the discharge height, the angle of inclination of the wetted sample carrier 6 'and optionally the discharge speed. To set the discharge height, the collecting device 34 and the pipetting unit 41 are aligned with the pipetting head 42 which holds the pipette with the pipette tip. In this case, a predetermined catchment area of the wetted sample carrier 6 'reaches the collecting position, the pipette tip is in the discharge position.
- the discharge height h a is preferably to a value in a range of 0, 1 to 800 mm, the inclination angle (double arrow c) of the wetted sample carrier 6 'to a value in a range of 0 ° to 90 °, the discharge speed to a value in Range from 0 to 10m / s.
- Step J A predetermined amount of the sample is dropped in one or more drops 8 from the dropping position to the predetermined catching area of the sample carrier 6 'to achieve bursting of the cell nuclei of the cells contained in the sample. It can be provided that the predetermined amount of the sample is dropped onto a cooled catchment area of the sample carrier 6 '.
- sample carrier 6 now carries at least one decomposed sample 9 and is therefore referred to below as sample carrier 6 "After ejection, a waiting period may be provided to achieve a bleeding of the cell components of the burst cells on the sample carrier, the sample carrier 6" having its angle of inclination can maintain.
- the sample carrier 6 can optionally be transferred to a parking position.
- Step K The sample carrier 6 "is guided in a first variant by means of the collecting device 34 to the drying device 51. There, the wetted and the zerplatze sample 9-bearing surface of the sample carrier 6" or the sample carrier 6 "is heated in total to prevent evaporation of the buffer and thus to achieve immobilization of the burst sample on the sample carrier 6 ".
- the angle of inclination of the sample support 6 can be set to a value in the range of 0 ° to 89 °, ie the angle of inclination can remain unchanged if it is not 90 °
- the heating can take place in a defined manner
- evaporation of the buffer and thus immobilization of the burst sample on the sample carrier 6 " can be achieved by a long waiting time in a defined climate in the working area.
- the drying by blower and / or existing in the housing 1 stream of the working gas from the climatic chamber 4 can be supported.
- Step L This step is optional.
- the sample carrier 6 "with the immobilized burst sample is now guided by means of the collecting device 34 to the analysis device 52.
- the analysis device 52 can be a microscope, for example in which the surface of the sample carrier 6 "carrying the fragmented sample 9 lies in a horizontal plane, where an automated, for example microscopic analysis of the immobilized decomposition can be carried out. burst sample 9 are performed.
- the sample carrier 6 " can be in a parking position.
- Step M The pipetting head 41 with the pipette 42, which carries the pipette tip used for receiving and discharging the sample, is guided to the collecting container 49. There, the pipette tip including the non-dropped portion of the sample is dropped. If a needle has been used instead of a pipette tip, then the needle is either washed, sterilized and reused in the device according to the invention, or discarded. In the latter variant, the needle can be washed externally and then sterilized.
- Step N The sample carrier 6 "with the immobilized burst sample is then moved back into the cooled storage area 2 by means of the collecting device 34. There, the sample carrier 6" can be deposited in the sample carrier receiving device 21.
- FIG. 4 illustrates a second process flow using a second embodiment of the device according to the invention.
- the second embodiment corresponds to the first embodiment except in the following points:
- the storage area 2 is not a refrigerated storage area.
- the storage area 2 has instead ambient temperature, ie room temperature.
- a heating / cooling plate 54 is instead provided as a temperature control device on which the sample carrier 6 'can lie.
- the tempering device may be part of the collecting device or formed separately.
- the holder for receiving the sample carrier which is also part of the collecting device, be designed so that it holds the sample carrier 6 'on the heating / cooling plate 54.
- the adjusting unit of the collecting device can then tilt the sample carrier 6 'into the predetermined angle of inclination with the heating / cooling plate 54.
- the tempering device has an adjusting unit for adjusting the angle of inclination of the sample carrier 6 'deposited on it, and that the temperature ration device is arranged in the device so that the sample carrier 6 'placed on it is in the catching position.
- the catching device does not need its own adjusting unit.
- the tool receiving devices 22, 23 and 24 are disposed in the cooled sub-work area 3a.
- Tracking means are provided for guiding aids from the tool receiving means 22, 23 and 24 into the wetting means 31.
- the pipette tip holder 47 holds two types of exchangeable tips or washable needles.
- the first type of tip or needles has a receiving volume of 200 ⁇
- the second type of tip or needles a receiving volume of 1000 ⁇ .
- the first grade is needed to mix the sample with the adjuvant added to adjust the given cell density in the sample container, the second grade to extract a defined amount of the sample from the sample container.
- FIG. 4 shows the non-tempered storage area 2 and the working area 3. It should be noted that the storage area 2 is shown twice in FIG. 4 because it is used to receive the empty sample carriers 6 'as well as the wetted ones and the sample carriers 6 "carrying the samples dropped off.
- The” Process flow "arrow illustrates the method sequence. The individual procedural steps are identified in the flowchart by capital letters which are located in a circle.
- the method sequence is explained on the basis of a sample and with reference to a sample carrier 6. However, it is also possible to use several samples simultaneously and / or several Sample carriers 6 are handled simultaneously, even if this is not expressly stated. In addition, several samples can be treated successively by means of the device according to the invention. For this, the treatment of a previous sample does not have to be completed. The time interval between two samples may depend on the waiting time and / or drying time needed to treat a sample.
- the auxiliaries may be the following auxiliaries: First auxiliaries: alcoholic solution as the first buffer component
- Second aid Acid solution as a second buffer component
- auxiliaries may be provided, for example one or more coloring agents, one or more reactive solutions and / or one or more washing solutions.
- the empty sample carriers 6 are uniquely identified by identification means (eg barcode).
- the containers in which the aids are located are clearly identified by means of identification (eg barcode).
- the storage area 2 has ambient temperature.
- a temperature of -20 ° C to room temperature In the rest of the work area, another defined climate is set, namely between -20 ° C and room temperature. It can be provided that all settings of the device as well as the process sequences, in particular the process parameters and sample information, are logged.
- the catcher 34 is shown with a double arrow to illustrate the movement of the catcher 34 in a horizontal direction.
- the double arrows d and e illustrate the movement of the catcher 34 in a vertical direction and another vertical direction.
- the collecting device 34 is therefore movable in the x, y and z directions.
- the movement of the pipetting unit 41 in a horizontal direction is shown by double arrow a.
- the pipetting head 42 is movable in the x, y and z directions.
- the inventive method starts when the sample carrier receiving device 21 filled with clean sample carriers and one or more samples, which are in sample containers in the inventive device on the sample supply 7 in the housing 1 in the sample-receiving device 48 in the sub-work area 3a are introduced.
- the samples may have previously been pretreated, for example centrifuged. They may also have been previously cooled to a temperature below room temperature.
- sample or samples are contained in sample containers that are uniquely identified by means of identification (eg barcode).
- the sample containers can be opened or closed. It can be provided that the sample containers are located in a rack which is introduced into the partial working area 3 a via the sample feed 7. It can be provided that several racks are introduced simultaneously into the partial work area 3a. It can be provided that the samples are further cooled after introduction into the sample receiving device 48, at least as long as they are in the sample containers.
- Step A The sample or samples introduced into the sample receiver 48 are identified by an identifier 50 (eg, bar code reader). This should allow a later assignment of the sample to a sample carrier 6.
- an identifier 50 eg, bar code reader
- Step B For each sample located in the sample receiver 48, the cell density of the sample is determined by the cell density meter. It can be provided that the cell density of the sample is adjusted when the measured cell density falls below or exceeds a predetermined value.
- one or more aids in defined amounts from the first, second and / or third auxiliary agent receiving device can be guided via the tracking means 53 to the sample container in which the sample is located.
- the aid (s) may be supplied in defined amounts from the first, second and / or third auxiliary receiving means to the sample container in which the sample is located by means of the first type of tips or needles the sample is dyed with a colorant and / or mixed with a reactive solution
- the colorant and reactive solution may be stored as auxiliary in tool receiving means in the cooled part work area 3a.
- Steps C and C2 The pipetting unit 41 is guided to the pipette tip holding device 47 and receives there one or more pipette tips or needles of the first or second kind.
- the pipette tips or needles of the first grade are used to mix samples with one or more tracked auxiliaries.
- the pipette tips or needles of the second grade are used for later recording of samples.
- Three variants are possible: In the first variant, the cell density measured in step B corresponded to the predetermined value. The pipetting unit first receives a pipette tip or needle of the second kind. In the second variant, the cell density measured in step B was above the predetermined value. The pipetting unit first picks up a pipette tip or needle of the first kind.
- predetermined amounts of one or more aids from the respective container of the aid receiving units 22, 23, 24 are guided into the sample in order to set a defined cell density.
- the supplied aids are mixed with the sample.
- the pipette tip or needle of the first type is then discarded, for example in collecting container 49.
- the pipetting unit 41 is again guided to the pipette tip holding device 47 and receives there a pipette tip or needle of the second kind.
- the cell density measured in step B was below the predetermined value.
- An alarm is triggered, which can be presented to the user via the computer 5.
- the sample is either not processed or treated according to the user default in response to the alarm.
- Step D The pipetting unit 41 is guided to the sample receiver 48.
- a pipette 46 of the pipetting unit 41 a defined amount of a sample is taken from a sample container. The sample enters the pipetting tip or needle of the pipette 46.
- Step E The catcher is moved to the storage area 2.
- a sample carrier 6 ' is removed from the sample carrier receiving device 21, wherein the sample carrier 6' enters the holding unit 35 and comes to lie there on the heating / cooling plate 54.
- the sample carrier 6 ' is then guided by means of the collecting device 34 to the wetting device 31. He arrives from the storage area 2 in the work area. 3
- Step F The sample carrier 6 'is guided by means of the collecting device to a second identification device 50 (eg barcode reader) and identified there. This should allow a later assignment of the sample to the sample carrier 6.
- a second identification device 50 eg barcode reader
- Step G The first and second aids are mixed to form an adjuvant mixture.
- the adjuvant mixture is the buffer.
- defined amounts of the first and second auxiliary means from the first and second auxiliary receiving means 22, 23 are guided via the guide means 33 to the wetting device 31 and mixed there to form a buffer.
- the defined amounts of the first and second aids may correspond to a fixed predetermined mixing ratio.
- step G is only required if no prefabricated buffer is used. In this case, the buffer from a tool-receiving device in which it is stored refrigerated, guided over the guide means 33 to the wetting device 31, where it can be readily used.
- the auxiliary mixture can be prepared and the samples can be wetted with drops of the auxiliary mixture. This eliminates the wetting device 31 and / or a mixing device.
- Step H The buffer is applied to the sample carrier 6 'via the nozzles 32.
- the sample carrier 6 ' is rinsed with the buffer and in particular the Surface of the sample carrier 6 'wetted with the buffer.
- Steps I and 12 According to the specifications of the user of the device according to the invention, the following parameters are now set: the discharge height, the angle of inclination of the wetted and resting on the heating / cooling plate 54 sample carrier 6 'and optionally the discharge speed. To set the discharge height, the collecting device 34 and the pipetting unit 41 are aligned with the pipetting head 42, which holds the pipette with the pipette tip.
- the discharge height h a is preferably to a value in a range of 0.1 to 800 mm, the inclination angle (double arrow c) of the wetted sample carrier 6 'to a value in a range of 0 ° to 90 °, the discharge speed to a value in Range from 0 to 10 m / s.
- the sample carrier is still located on the heating / cooling plate.
- Step J A predetermined amount of the sample is dropped in one or more drops 8 from the dropping position to the predetermined catching area of the sample carrier 6 'to achieve bursting of the cell nuclei of the cells contained in the sample.
- the sample carrier now carries at least one decomposed sample 9 and is therefore referred to below as sample carrier 6 "
- the sample carrier is optionally in a cooled state during the dropping of the sample, whereby it can either be actively cooled or positioned on a cold block or with cold buffer To coat the sample carrier with cold buffer, the sample carrier can be immersed, for example, in the buffer.
- Step K The sample carrier 6 "is guided into a drying area by means of the collecting device 34. There, the angle of inclination of the sample carrier 6" becomes 0 ° in which the surface of the sample carrier 6 "bearing the fragmented sample 9 is horizontal Subsequently, the sample carrier 6 "carrying the wetted and disintegrated sample 9 is heated to a predetermined temperature for a predetermined period of time by means of the heating / cooling plate 54 in order to prevent evaporation of the buffer and thus immobilization of the buffer.
- the temperature may, for example, be 90 ° C., the time period about 3 to 4 minutes, optionally the drying by means of blowers can be assisted 6 'or 6 "may be formed in one position but also in separate positions.
- Step L This step is optional.
- the sample carrier 6 "with the immobilized burst sample is now guided by means of the collecting device 34 to the analysis device 52.
- the analysis device 52 can be a microscope, for example In this case, the sample carrier 6 "can be in a parked position.
- Step M The pipetting head 41 with the pipette 42, which carries the second type pipette tip used for receiving and ejecting the sample, is guided to the collecting container 49. There, the pipette tip is thrown off including the part of the sample that has not been removed. If a needle has been inserted instead of a pipette tip, the needle is thrown off. The needle can then be washed and then sterilized. If the needle is not thrown off, it can possibly be washed, sterilized and reused in the device according to the invention.
- Step N The sample carrier 6 "with the immobilized burst sample is now moved back into the storage area 2 by means of the collecting device 34. There, the sample carrier 6" can be deposited in the sample carrier receiving device 21. LIST OF REFERENCE NUMBERS
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Thermal Sciences (AREA)
- Analytical Chemistry (AREA)
- Mechanical Engineering (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Sampling And Sample Adjustment (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102016120726.8A DE102016120726A1 (de) | 2016-10-28 | 2016-10-28 | Vorrichtung und Verfahren zur Behandlung biologische Zellen enthaltender Proben, insbesondere von Blut- oder Zellproben |
PCT/DE2017/100923 WO2018077352A1 (de) | 2016-10-28 | 2017-10-28 | Vorrichtung und verfahren zur behandlung biologische zellen enthaltender proben |
Publications (1)
Publication Number | Publication Date |
---|---|
EP3532599A1 true EP3532599A1 (de) | 2019-09-04 |
Family
ID=60381987
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP17800378.6A Pending EP3532599A1 (de) | 2016-10-28 | 2017-10-28 | Vorrichtung und verfahren zur behandlung biologische zellen enthaltender proben |
Country Status (4)
Country | Link |
---|---|
US (2) | US10907129B2 (de) |
EP (1) | EP3532599A1 (de) |
DE (1) | DE102016120726A1 (de) |
WO (1) | WO2018077352A1 (de) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102016120726A1 (de) * | 2016-10-28 | 2018-05-03 | Als Automated Lab Solutions Gmbh | Vorrichtung und Verfahren zur Behandlung biologische Zellen enthaltender Proben, insbesondere von Blut- oder Zellproben |
BR112020004016A2 (pt) * | 2017-08-31 | 2020-09-08 | Monsanto Technology Llc | sistema de entrega de sementes |
DE102018111064A1 (de) * | 2018-05-08 | 2019-11-14 | Lockcon Ag | Öffnungsmaschinenvorrichtung und Probenbehälter zur Verwendung mit der Öffnungsmaschinenvorrichtung |
CN114184438B (zh) * | 2020-09-14 | 2024-05-10 | 长春理工大学 | 一种提取染色体的装置和方法 |
Family Cites Families (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6850252B1 (en) * | 1999-10-05 | 2005-02-01 | Steven M. Hoffberg | Intelligent electronic appliance system and method |
EP0633207A1 (de) * | 1993-07-07 | 1995-01-11 | Siemens Aktiengesellschaft | Transportsystem zum Transport von Proben zu unterschiedlichen Behandlungseinrichtungen |
US5891734A (en) * | 1994-08-01 | 1999-04-06 | Abbott Laboratories | Method for performing automated analysis |
US6122396A (en) * | 1996-12-16 | 2000-09-19 | Bio-Tech Imaging, Inc. | Method of and apparatus for automating detection of microorganisms |
CA2342798A1 (en) * | 1998-09-02 | 2000-03-16 | Langerhans Aps | Apparatus for isolation of particles, preferably cell clusters |
DE10135091A1 (de) * | 2001-07-15 | 2003-01-30 | Universitaetsklinikum Charite | Verfahren, Einrichtung und Objektträger zur Mikrodissektion |
WO2003048705A1 (en) * | 2001-12-05 | 2003-06-12 | The Regents Of The University Of California | Robotic microscopy systems |
US7776584B2 (en) * | 2003-08-01 | 2010-08-17 | Genetix Limited | Animal cell colony picking apparatus and method |
US7783383B2 (en) * | 2004-12-22 | 2010-08-24 | Intelligent Hospital Systems Ltd. | Automated pharmacy admixture system (APAS) |
US8457781B2 (en) * | 2007-09-13 | 2013-06-04 | Lockheed Martin Corporation | Facility wide mixed mail sorting and/or sequencing system and components and methods thereof |
WO2010144859A2 (en) * | 2009-06-12 | 2010-12-16 | Life Technologies Corporation | Devices and methods for automated sample preparation, assay, and analysis |
US8943785B2 (en) * | 2009-08-20 | 2015-02-03 | Pioneer Hi Bred International Inc | Automated high-throughput seed processing apparatus |
DE102010003223B4 (de) * | 2010-03-24 | 2014-09-18 | Albert-Ludwigs-Universität Freiburg | Vorrichtung zum Einsetzen in einen Rotor einer Zentrifuge, Zentrifuge und Verfahren zum fluidischen Koppeln von Kavitäten |
CN202297580U (zh) * | 2011-11-01 | 2012-07-04 | 浙江世纪康大医疗科技有限公司 | 一种提高生化试剂和生物酶效能装置 |
DE102016120726A1 (de) * | 2016-10-28 | 2018-05-03 | Als Automated Lab Solutions Gmbh | Vorrichtung und Verfahren zur Behandlung biologische Zellen enthaltender Proben, insbesondere von Blut- oder Zellproben |
US20190259108A1 (en) * | 2018-02-20 | 2019-08-22 | Osram Gmbh | Controlled Agricultural Systems and Methods of Managing Agricultural Systems |
-
2016
- 2016-10-28 DE DE102016120726.8A patent/DE102016120726A1/de active Pending
-
2017
- 2017-10-28 WO PCT/DE2017/100923 patent/WO2018077352A1/de unknown
- 2017-10-28 EP EP17800378.6A patent/EP3532599A1/de active Pending
-
2019
- 2019-04-28 US US16/396,762 patent/US10907129B2/en active Active
-
2020
- 2020-12-23 US US17/133,589 patent/US20210115387A1/en active Pending
Also Published As
Publication number | Publication date |
---|---|
WO2018077352A1 (de) | 2018-05-03 |
US20210115387A1 (en) | 2021-04-22 |
DE102016120726A1 (de) | 2018-05-03 |
US10907129B2 (en) | 2021-02-02 |
US20190249133A1 (en) | 2019-08-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2018077352A1 (de) | Vorrichtung und verfahren zur behandlung biologische zellen enthaltender proben | |
DE102008054066B4 (de) | Verfahren zum Bearbeiten von Gewebeproben unter Verwendung eines Sensors | |
DE19652784A1 (de) | Vorrichtung (Küvette) zur Aufnahme und Speicherung von Flüssigkeiten und zur Durchführung optischer Messungen | |
EP1664727A1 (de) | Vorrichtung und verfahren zur automatisierten durchführung v on laborarbeitsschritten | |
EP3039399B1 (de) | Lasermikrodissektionssystem und lasermikrodissektionsverfahren | |
EP2623202B1 (de) | Verfahren zum erwärmen eines flüssigkeitsvolumens in einer beheizten pipettiernadel, pipettiervorrichtung und automatisches analysegerät | |
WO2014009066A1 (de) | Verfahren und analysevorrichtung zur mikroskopischen untersuchung eines gewebeschnittes oder eines zellausstrichs | |
EP2936144B1 (de) | Verfahren und vorrichtung zur durchführung einer dünnschichtchromatographie | |
WO2015032630A1 (de) | Mikrodissektionsgerät und verfahren zum isolieren von zellen eines vorbestimmten zelltyps | |
EP2646835B1 (de) | Verfahren und vorrichtung zur automatischen erfassung von substanzen für analysen | |
DE69030957T2 (de) | Probentransportsystem für optisches überwachungssystem | |
DE102012013680A1 (de) | Vorrichtung sowie Verfahren zur Inkubation von Patientenproben | |
EP3130926A1 (de) | Pipettiervorrichtung mit zwei temperatursensoren | |
WO2010112016A1 (de) | Verfahren und vorrichtung zur behandlung von trägerfixiertem material | |
DE4041411A1 (de) | Chromatographieverfahren zum analysieren biologischer proben und mit einem solchen verfahren arbeitender fluessigphasenchromatographie-anlaysator | |
DE102008056584B4 (de) | Verfahren und Vorrichtung zur Behandlung von Objekten | |
EP0643989B1 (de) | Verfahren und System zur Mischung von Flüssigkeiten | |
EP2710109B1 (de) | Verfahren und verwendung einer vorrichtung zur automatisierten positionsbestimmung eines mikrosystems zur manipulation eines sphärischen mikro-objekts | |
EP1594613B1 (de) | Verfahren zur untersuchung zellulärer proben | |
WO2013064237A2 (de) | Automatische strukturbestimmung | |
EP1833601A1 (de) | Verfahren und vorrichtung zur entnahme und analyse von proben | |
EP3334531B1 (de) | Vorrichtung und verfahren zur bestimmung der migrationsfähigkeit amöboid beweglicher zellen | |
EP3173794B1 (de) | Verfahren zum transfer eines flüssigkeitsvolumens in einem analysegerät | |
WO1991012516A1 (de) | Vorrichtung und verfahren zur bestimmung klinisch-chemischer parameter u.a. | |
WO2021213635A1 (de) | Pipettiervorrichtung und verfahren zur bearbeitung einer fluiden probe |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: UNKNOWN |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE |
|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE |
|
17P | Request for examination filed |
Effective date: 20190520 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
AX | Request for extension of the european patent |
Extension state: BA ME |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: UNGLAUB, DAVID Inventor name: EBERHARDT, JENS Inventor name: OPEL, MATTHIAS Inventor name: BORNMANN, GERD |
|
DAV | Request for validation of the european patent (deleted) | ||
DAX | Request for extension of the european patent (deleted) | ||
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: SARTORIUS AUTOMATED LAB SOLUTIONS GMBH |