Apparatus for processing samples containing biological cells

Abstract

An apparatus and a method for the automated processing of samples containing biological cellsin particular, of blood samples or other cell samples is provided. The apparatus has a sample receiving device configured to receive samples, an auxiliary material receiving device configured to receive auxiliary materials, a sample carrier receiving device configured to receive sample carriers, a discharge device configured to discharge samples from a discharge position, and a capture device configured to capture discharged samples in a capture position to obtain prepared samples, and at least one temperature control device for controlling the temperature of at least one of the sample carriers. The distance between the discharge position and the capture position can be adjusted to a prespecified height. The method includes wetting a temperature-controlled sample carrier with at least one auxiliary material and discharging the sample onto the wetted temperature-controlled sample carrier from a prespecified height.

Claims

1. An apparatus for the automated processing of a sample containing biological cells, the apparatus comprising: a temperature-controlled storage area; a working area; a sample receiving device configured to receive the sample; an auxiliary material receiving device configured to receive auxiliary materials; a sample carrier receiving device configured to receive sample carriers; a discharge device configured to discharge the sample from a discharge position; at least one temperature control device configured to control a temperature of at least one of the sample carriers, and a capture device configured to capture the sample discharged from the discharge position in a capture position to obtain a prepared sample, and a distance between the discharge position and the capture position being adjusted to a prespecified height of between 0.1 to 800 mm, the capture device having a holder configured to receive at least one of the sample carriers, said holder being configured such that the at least one sample carrier is arranged in the capture position to obtain the discharged sample.

2. The apparatus according to claim 1, wherein the sample containing the biological cells is a blood sample or another cell sample.

3. The apparatus according to claim 1, further comprising a housing in which components of the apparatus are arranged, wherein in the housing, physical and/or chemical conditions are set which differ from the conditions of the environment.

4. The apparatus according to claim 1, wherein the discharge device includes a pipetting unit with a pipetting head movable at least along one axis.

5. The apparatus according to claim 1, wherein the temperature-controlled storage area includes at least one of the auxiliary material receiving device and the sample carrier receiving device arranged therein.

6. The apparatus according to claim 1, wherein the working area includes at least the discharge device and the capture device arranged therein.

7. The apparatus according to claim 1, wherein: the temperature controlled sample receiving device is arranged in the working area or in the storage area.

8. The apparatus according to claim 1, further comprising a wetting device configured to wet the sample carriers with at least one of the auxiliary materials.

9. The apparatus according to claim 1, further comprising an analysis unit configured to analyze the prepared sample.

10. The apparatus according to claim 1, wherein: the capture device has an actuator configured to adjust a prespecified angle of inclination of the at least one sample carrier, and the angle of inclination of the at least one sample carrier is adjustable to a value between 0 and 90 relative to a horizontal plane.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The disclosure will now be described with reference to the drawings wherein:

(2) FIG. 1 shows a top view of an apparatus according to a first exemplary embodiment of the disclosure;

(3) FIG. 2 shows a schematic representation of the apparatus according to the first exemplary embodiment of disclosure, without a housing;

(4) FIG. 3 shows a first flow diagram illustrating a first method sequence provided for the apparatus according to the first exemplary embodiment of the disclosure; and

(5) FIG. 4 shows a second flowchart illustrating a second method sequence provided for the apparatus according to the second exemplary embodiment of the disclosure.

DESCRIPTION OF EXEMPLARY EMBODIMENTS

(6) The first exemplary embodiment of the apparatus, shown in FIGS. 1 and 2, has a housing 1. In the housing 1, a cooled storage area 2, and a working area 3 are formed as a storage area. The storage area 2 and the working area 3 are chambers within the housing 1, which are connected to each other via an opening or a lock. A climatic chamber 4 in which there are means for preparing and a conveyor configured to convey a working gas are arranged in the housing above the cooled storage area 2 and the working area 3. The working gas is the gaseous medium in the housingfor example, air. The working gas can be prepared, for example filtered, using the means for preparing the working gas. The means for preparing the working gas can be, for example, a filter. Using the conveyor configured to convey the working gas, the working gas can be conveyed from the cooled storage area 2, the working area 3, or both into the climatic chamber 4 to the means for preparing the working gas, and prepared there. Using further conveyors configured to convey the working gas, the prepared working gas can then be returned to the cooled storage area 2, the working area 3, or both. A data processing device 5for example, a computeris arranged outside the housing 1, and serves to control the components of the apparatus arranged in the housing. In the housing there is a sample feed 7 (see FIG. 3) which allows for the introduction of samples into the working area 3.

(7) The cooled storage area 2 has a temperature of from 20 C. to room temperature. The working area 3 has a temperature of from 20 C. to room temperature.

(8) A sample carrier receiving device 21 is arranged in the cooled storage area 2, enabling the reception of a plurality of sample carriers 6. The sample carrier receiving device 21 can be a rack. Furthermore, in the cooled storage area 2, there are a first auxiliary material receiving device 22 configured to receive a first auxiliary material, and a second auxiliary material receiving device 23 configured to receive a second auxiliary material. The first auxiliary material receiving device 22 and the second auxiliary material receiving device 23 can be containers. The first auxiliary material can be a first substance needed to make a buffer. The first substance can be an alcohol. The second auxiliary material can be a second substance needed to make a buffer. The second substance can be an acid. In addition, a third auxiliary material receiving device 24 (see FIG. 3) can be arranged in the cooled storage area 2 to receive a third auxiliary material. The third auxiliary material receiving device 24 can be a container. The third auxiliary material can be, for example, a dye or a reactive solution. Further auxiliary material receiving devices can be arranged in the cooled storage area 2. It should be expressly understood that it is not absolutely necessary to produce a buffer in the apparatus according to the disclosurefor example, from the first auxiliary material and the second auxiliary material. Rather, the buffer can be introduced in ready-to-use form in an auxiliary material receiving device into the cooled storage area 2.

(9) The samples are expediently held in sample containersfor example, in test tubes. The auxiliary materials are expediently in auxiliary material containers. The sample containers, sample carriers 6 and/or the auxiliary material containers can be identified with identifiers (for example, a barcode), enabling a unique identification of each individual sample, each sample carrier and, if applicable, each individual auxiliary material. This permits the assignment and tracking of samples, sample carriers 6, and auxiliary materials. The identifiers (for example, a barcode) can be, for example, optical codesin particular, a barcodeor RFID tags or other identifiers. The identifiers (for example, barcodes) can be applied by known methods to the sample containers, the sample carriers 6, and the auxiliary material containers. The identifiers make it possible to comprehensively document the individual method steps together with the process parametersfor example, in reports.

(10) In the working area 3, there is a wetting device 31 configured to wet sample carriers 6. The wetting device 31 has a mixing device (not shown) for producing an auxiliary material mixture. The wetting device 31 also has nozzles 32, via which the auxiliary material mixture can be applied to the sample carrier 6. The nozzles 32 can 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. Conveyors 33for example, hoses and a pumpare provided to convey the auxiliary materials from the first and second auxiliary material receiving devices 22, 23 to the mixing device. As an alternative to the pump, a pump system can be used. It should be expressly understood that it is not absolutely necessary to produce a buffer in the apparatus according to an aspect of the disclosurefor example, from the first auxiliary material and the second auxiliary material. Rather, the buffer can be introduced in ready-to-use form in an auxiliary material receiving device into the cooled storage area 2. Thus, no mixing device is required. Rather, the buffer can be conveyed via a conveyor from the auxiliary material receiving device in which the ready-to-use buffer is held to a wetting device which has no mixing device.

(11) By the wetting device 31, an auxiliary material or a mixture of auxiliary materials can be applied to a sample carrier 6 in a defined mixing ratio. A collecting unit 40for example, a bowlis arranged below the wetting device 31, and receives the portion of the auxiliary material or the auxiliary material mixture that does not adhere to the sample carrier 6.

(12) A capture device 34 is provided for removing the sample carrier 6 from the sample carrier receiving device 21. The capture device 34 has a holding unit 35 for the sample carrier 6 and an actuator 36 for inclining the sample carrier 6 located in the holding unit 35. The holding unit 35 is movable in the x-, y-, and z-directions. The holding unit 35 and the actuator 36 may be attached via an arm 37 to a runner 38 which runs on a rail 39. The runner 38 can be moved on the rail 39for example by a servomotorin a horizontal direction (x-coordinate). The rail 39 is mounted (not shown) in the housing 1 in such a manner that the rail 39 is movable in the vertical direction (z-coordinate), and a second horizontal direction (y-coordinate).

(13) In addition, 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-, and z-directions. For this purpose, the pipetting head 42 can be attached via an arm 43 to a runner 44 which runs on a rail 45. The runner 44 can be moved on the rail 45for example, by a servomotorin a horizontal direction (x-coordinate). The rail 45 is mounted (not shown) in the housing 1 in such a manner that the rail 45 is movable in the vertical direction (z-coordinate), and a second horizontal direction (y-coordinate). The pipetting head can hold one or more pipettes 46. The pipettes can have pipette tips or needles. The pipette tips of the pipettes 46 are exchangeable. The pipette needles of the pipettes 46 are exchangeable. The pipette needles can be washable.

(14) In the working area 3, there is also a pipette tip holding device 47 which can receive one or more pipette tips or needles. Furthermore, a sample receiving device 48 can be arranged in the working area 3, and can receive one or more samples. The sample receiving device 48 can be temperature-controlledtypically cooled. The sample receiving device 48 can include a cell density measuring unit 49 with which it is possible to determine the cell density of the samples prior to the discharging. In addition, a collecting vessel 49 for used pipette tips or needles can be arranged in the working area 3. The collecting vessel 49 can include an ejecting mechanism to separate the pipette tips or needles from a pipette.

(15) One or more identification devices 50 (for example, bar code readers) can be arranged in the working area 3, allowing identification of identifiers (for example, barcodes). In addition, a drying device 51for example, a heating devicewhich enables drying and thus immobilization of the sample discharged onto the sample carrier can be arranged in the working area 3. In addition, an analysis device 52for example, a microscopecan be arranged in the working area 3, which allows an analysis of the sample which has been discharged onto the sample carrier and, if applicable, dried.

(16) The method according to an aspect of the disclosure will now be explained in more detail in one exemplary embodiment with reference to FIG. 3. The flowchart shown in FIG. 3 shows the cooled storage area 2 and the working area 3. It should be noted that the cooled storage area 2 is shown twice in FIG. 3, because it serves both for receiving the empty sample carriers 6 and the wetted sample carriers 6 carrying the discharged samples. The process flow arrow illustrates the process flow. The individual process steps are identified in the flowchart by capital letters in a circle.

(17) The method sequence is explained using a sample and a sample carrier 6 as an example. However, it is also possible to simultaneously handle a plurality of samples and/or a plurality of sample carriers 6, even if this is not expressly stated. In addition, several samples can be handled successively by the apparatus. For this, the processing of a previous sample does not have to be completed. The time interval between two samples can depend on the waiting time and/or drying time needed to process a sample.

(18) In the initial state of the apparatus according to an aspect of the disclosure, empty sample carriers 6 are located in the sample carrier receiving device 21, and the auxiliary materials required for producing the bufferin particular, the first and second auxiliary materialsare located in the first and second auxiliary material receiving devices 22, 23, respectively. In addition, pipette tips or needles are in the pipette tip holding device 47. The auxiliary materials may be the follow auxiliary materials:

(19) First auxiliary material: alcohol solution as the first buffer component.

(20) Second auxiliary material: acid solution as a second buffer component.

(21) Third auxiliary material: dye.

(22) Fourth auxiliary material: reactive solution.

(23) Fifth auxiliary material: washing solution.

(24) Further auxiliary materialsfor example, further dyes, reactive solutions and/or washing solutionscan be included.

(25) The empty sample carriers 6 are uniquely identified by identifiers (for example, a barcode). The containers in which the auxiliary materials are located are clearly identified by identifiers (for example, a barcode). The cooled storage area 2 is cooled to a temperature of from 20 C. to room temperature. In the working area, a defined climate is set, namely from 20 C. to room temperature.

(26) All settings of the apparatus, as well as the method processesin particular, the process parameters and sample informationcan be logged.

(27) In FIG. 3, the capture device 34 is shown with a double arrow to illustrate the movement of the capture device 34 in a horizontal direction. The double arrows d and e illustrate the movement of the capture device 34 in a vertical direction and a further horizontal direction. The capture device 34 is thus 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 movement of the pipetting unit 41 in the vertical direction is shown by double arrow b. The pipetting head 42 is movable in the x-, y- and z-directions.

(28) The method starts when the sample carrier receiving device 21 filled with clean sample carriers and one or more samples positioned in sample containers are introduced into the apparatus via the sample feed 7 in the housing 1, into the sample receiving device 48 and into the working area 3. The samples may have previously been pre-processedfor example, centrifuged. They can also have been previously cooled to a temperature below room temperature. The sample and/or the samples are located in sample containers which are uniquely identified by identifiers (for example, a barcode). The sample containers can be opened or closed. The sample containers can be positioned in a rack which is introduced into the working area 3 via the sample feed 7. Several racks can be introduced simultaneously into the working area 3. The samples can be further cooled after introduction into the sample receiving device 48at least as long as they are in the sample containers.

(29) Step A: The sample and/or the samples introduced into the sample receiving device 48 are identified by an identification device 50 (for example, a bar code reader). This is intended to allow a later assignment of the sample to a sample carrier 6.

(30) Step B: This step is optional. For each sample located in the sample receiving device 48, the cell density of the sample is determined by the cell density measuring device. The cell density of the sample can be adjusted if the measured cell density falls below or exceeds a prespecified value. This process can be referred to as revising the cell density. The cell density can be determined, for example, by an optical sensor and adjusted by the pipetting system and the buffer. The sample can be stained with a dye and/or mixed with a reactive solution. The dye and the reactive solution can be stored as auxiliary materials in auxiliary material receiving devices in the cooled storage area 2.

(31) Step C: The pipetting unit 41 is conveyed to the pipette tip holding device 47 where it receives one or more pipette tips or needles. The pipette tips or needles are used to take up samples later.

(32) Step D: The pipetting unit 41 is conveyed to the sample receiving device 48. A defined amount of a sample is taken from a sample container by a pipette 46 of the pipetting unit 41. The sample enters the pipette tip or needle of the pipette 46.

(33) Step E: The capture device is moved into the cooled storage area 2. By the capture device 34, a sample carrier 6 is removed from the sample carrier receiving device 21, and the sample carrier 6 is moved into the holding unit 35. The sample carrier 6 is then conveyed by the capture device 34 to the wetting device 31. In the process, it passes from the cooled storage area 2 into the working area 3.

(34) Step F: The sample carrier 6 is conveyed by the capture device to a second identification device 50 (for example, a barcode reader), and identified there. The purpose of this is to allow a later assignment of the sample to the sample carrier 6.

(35) Step G: The first and second auxiliary materials are mixed to form an auxiliary material mixture. The auxiliary material mixture is the buffer. For the preparation of the auxiliary material mixture, defined amounts of the first and second auxiliary materials from the first and second auxiliary material receiving device 22, 23 are conveyed via the conveyor 33 to the wetting device 31 and mixed there to form a buffer. The defined amounts of the first and second auxiliary materials can correspond to a prespecified mixing ratio. If step B is included in the method, the cell density of the sample can be taken into account when the defined amount is specified. It should be noted that step G is only required if no prefabricated buffer is used. In this case, the buffer is conveyed from an auxiliary material receiving device in which it is stored and cooled, via the conveyor 33, to the wetting device 31, where it can be used directly. It should also be noted that alternatively, the auxiliary material mixture can be prepared and the samples can be wetted with drops of the auxiliary material mixture by the pipetting system. This eliminates the wetting device 31, a mixing device, or both.

(36) Step H: The buffer is applied to the sample carrier 6 via the nozzles 32. In this case, the sample carrier 6 is rinsed with the buffer, and particularly 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.

(37) Step I: According to the specifications of the user of the apparatus, 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 capture device 34 and the pipetting unit 41with the pipetting head 42 which holds the pipette with the pipette tipare aligned relative to each other. In this case, a prespecified capture region of the wetted sample carrier 6 moves into the capture position, and the pipette tip moves into the discharge position. The discharge height ha is typically set to a value in a range from 0.1 to 800 mm; the angle of inclination (double arrow c) of the wetted sample carrier 6 is set to a value in a range from 0 to 90; and the discharge speed is set to a value in the range from 0 to 10 m/s.

(38) Step J: A prespecified amount of the sample is discharged in one or more drops 8 from the discharge position to the prespecified capture region of the sample carrier 6, to achieve bursting of the cell nuclei of the cells contained in the sample. The prespecified amount of the sample can be discharged onto a cooled capture region of the sample carrier 6. The sample carrier now carries at least one burst sample 9 and is therefore hereinafter referred to as the sample carrier 6. After the discharge, a waiting time can be included in the method to achieve running of the cell components of the broken cells on the sample carrier, wherein the sample carrier 6 can maintain its angle of inclination. The sample carrier 6 may optionally be transferred to a parked position.

(39) Step K: The sample carrier 6 is conveyed in a first variant by the capture device 34 to the drying device 51. There, the surface of the sample carrier 6 or the sample carrier 6 which carries the wetted and burst sample 9 is heated as a whole to achieve evaporation of the buffer, and thus immobilization of the burst sample on the sample carrier 6. The angle of inclination of the sample carrier 6 can be set to a value in the range from 0 to 89that is, the angle of inclination can remain unchanged if it is not 90. The heating can be done in a defined manner. This may involve specifying a drying temperature and specifying a drying duration. In a second variant, 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. Optionally, the drying can be supported by a blower and/or a flow of the working gas from the climatic chamber 4, already flowing in the housing 1.

(40) Step L: This step is optional. The sample carrier 6 with the immobilized, burst sample is now conveyed by the capture device 34 to the analysis device 52. The analysis device 52 can be, for example, a microscope. The angle of inclination of the sample carrier 6 can be changedfor example, to an inclination angle of 0, in which the surface of the sample carrier 6 carrying the fragmented sample 9 lies in a horizontal plane. There, an automatedfor example, microscopicanalysis of the immobilized burst sample 9 can be carried out. In this case, the sample carrier 6 can be in a parked position.

(41) Step M: The pipetting head 41 with the pipette 42 which carries the pipette tip used for taking up and discharging the sample is conveyed to the collecting container 49. There, the pipette tip including the non-discharged portion of the sample is discarded. If a needle has been used instead of a pipette tip, the needle is either washed, sterilized and reused in the apparatus, or is discarded. In the latter variant, the needle can be washed externally and then sterilized.

(42) Step N: The sample carrier 6 with the immobilized, burst sample is now moved back into the cooled storage area 2 by the capture device 34. There, the sample carrier 6 can be stored in the sample carrier receiving device 21.

(43) The flowchart shown in FIG. 4 illustrates a second method sequence using a second exemplary embodiment of the apparatus according to the disclosure. The second exemplary embodiment corresponds to the first embodiment except in the following points:

(44) (i) The storage area 2 is not a cooled storage area. The storage area 2 is instead at ambient temperaturethat is, room temperature. Rather, for controlling the temperature of the sample carrier, there is a heating/cooling plate 54 as the temperature control device, on which the sample carrier 6 can be placed. The temperature control device can be part of the capture device, or formed separately. For example, the holder for receiving the sample carrier, which is also part of the capture device, can be configured in such a manner that it holds the sample carrier 6 on the heating/cooling plate 54. The actuator of the capture device can then tilt the sample carrier 6 into the prespecified angle of inclination with the heating/cooling plate 54. In a separate configuration of the temperature control device, the temperature control device can have an actuator for adjusting the angle of inclination of the sample carrier 6 deposited on it, and the temperature control device can be arranged in the apparatus in such a manner that the sample carrier 6 deposited on it is in the capture position. In this case, the capture device does not need its own actuator.

(45) (ii) A cooled working area sub-region is constructed in the working area 3. The cooled working area sub-region 3a has a temperature of from 20 C. to room temperature; the remainder of the working area 3 has a different temperature, from 20 C. to room temperature.

(46) (iii) The auxiliary material receiving devices 22, 23 and 24 are arranged in the cooled working area sub-region 3a.

(47) (iv) Supply means are provided for conveying auxiliary materials from the auxiliary material receiving devices 22, 23 and 24 into the wetting means 31.

(48) (v) The pipette tip holding device 47 holds two types of exchangeable tips or washable needles. In one exemplary embodiment, the first type of tips or needles has an uptake volume of 200 l, and the second type of tips or needles has an uptake volume of 1000 l. The first type is needed for mixing the sample with the auxiliary material in the sample container, which is added to adjust the given cell density. The second type is needed to extract a defined amount of the sample from the sample container.

(49) (vi) No drying device is included.

(50) The flowchart shown in FIG. 4 shows the storage area 2 and the working area 3, which are not temperature-controlled. It should be noted that the non-temperature-controlled storage area 2 is shown twice in FIG. 4, because it serves both for receiving the empty sample carriers 6 and the wetted sample carriers 6 carrying the discharged samples. The process flow arrow illustrates the process flow. The individual process steps are identified in the flowchart by capital letters in a circle.

(51) The method sequence is explained using a sample and a sample carrier 6 as an example. However, it is also possible to simultaneously handle a plurality of samples and/or a plurality of sample carriers 6, even if this is not expressly stated. In addition, several samples can be handled successively by the apparatus. For this, the processing of a previous sample does not have to be completed. The time interval between two samples can depend on the waiting time and/or drying time needed to process a sample.

(52) In the initial state of the apparatus, empty sample carriers 6 are located in the sample carrier receiving device 21, and the auxiliary materials required to produce the bufferin particular, the first and second auxiliary materialsare located in the first and second auxiliary material receiving devices 22, 23, respectively. In addition, pipette tips or needles are in the pipette tip holding device 47. The auxiliary materials may be the follow auxiliary materials:

(53) First auxiliary material: alcohol solution as the first buffer component.

(54) Second auxiliary material: acid solution as a second buffer component.

(55) Third auxiliary material: dye or reactive solution.

(56) Further auxiliary materialsfor example, one or more dyes, one or more reactive solutions, and/or one or more washing solutionscan be included.

(57) The empty sample carriers 6 are uniquely identified by identifiers (for example, a barcode). The containers in which the auxiliary materials are located are clearly identified by identifiers (for example, a barcode). The storage area 2 is at ambient temperature. The temperature of the working area sub-region 3a is set to from 20 C. to room temperature. In the rest of the working area, a different defined climate is setnamely, between 20 C. and room temperature.

(58) All settings of the apparatus, as well as the method processesin particular, the process parameters and sample informationcan be logged.

(59) In FIG. 4, the capture device 34 is shown with a double arrow to illustrate the movement of the capture device 34 in a horizontal direction. The double arrows d and e illustrate the movement of the capture device 34 in a vertical direction and a further vertical direction. The capture device 34 is thus 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 movement of the pipetting unit 41 in the vertical direction is shown by double arrow b. The pipetting head 42 is movable in the x-, y- and z-directions.

(60) The method starts when the sample carrier receiving device 21 filled with clean sample carriers and one or more samples positioned in sample containers are introduced into the apparatus via the sample feed 7 in the housing 1, into the sample receiving device 48 and into the working area sub-region 3a. The samples may have previously been pre-processedfor example, centrifuged. They can also have been previously cooled to a temperature below room temperature. The sample and/or the samples are located in sample containers which are uniquely identified by identifiers (for example, a barcode). The sample containers can be opened or closed. The sample containers can be positioned in a rack which is introduced into the working area sub-region 3a via the sample feed 7. Several racks can be introduced simultaneously into the working area sub-region 3a. The samples can be further cooled after introduction into the sample receiving device 48at least as long as they are in the sample containers.

(61) Step A: The sample and/or the samples introduced into the sample receiving device 48 are identified by an identification device 50 (for example, a bar code reader). This is intended to allow a later assignment of the sample to a sample carrier 6.

(62) Step B: For each sample located in the sample receiving device 48, the cell density of the sample is determined by the cell density measuring device. The cell density of the sample can be adjusted if the measured cell density falls below or exceeds a prespecified value. This process can be referred to as revising the cell density. For this purpose, one or more auxiliary materials can be conveyed in defined amounts via the supply means 53 from the first, second and/or third auxiliary material receiving device(s) to the sample container in which the sample is located. Alternatively, the auxiliary material(s) can be conveyed in defined amounts to the sample container in which the sample is situated, by the first type of tips or needles in step C2, from the first, second and/or third auxiliary material receiving device(s). The sample can be stained with a dye and/or mixed with a reactive solution. The dye and the reactive solution can be stored as auxiliary materials in auxiliary material receiving devices in the cooled working area sub-region 3a.

(63) Steps C and C2: The pipetting unit 41 is conveyed to the pipette tip holding device 47 and receives there one or more pipette tips or needles of the first or second type. The pipette tips or needles of the first type are used to mix samples with one or more supplied auxiliary materials. The pipette tips or needles of the second type are used later for taking up samples. Three variants are possible: In the first variant, the cell density measured in step B corresponded to the prespecified value. The pipetting unit first takes a pipette tip or needle of the second type. In the second variant, the cell density measured in step B was above the prespecified value. The pipetting unit first takes a pipette tip or needle of the first type. Subsequently, by the pipetting unit or the supply means, prespecified amounts of one or more auxiliary material(s) are conveyed from the respective container of the auxiliary material receiving units 22, 23, 24 into the sample to reach a defined cell density. The supplied auxiliary materials are mixed with the sample. The pipette tip or needle of the first type is then discardedfor example, into the collecting vessel 49. Then, the pipetting unit 41 is conveyed again to the pipette tip holding device 47, where it takes a pipette tip or needle of the second type. In the third variant, the cell density measured in step B was below the prespecified value. An alarm is triggered, and can be presented to the user via the computer 5. The sample is either not processed, or is processed according to the user default in response to the alarm.

(64) Step D: The pipetting unit 41 is conveyed to the sample receiving device 48. A defined amount of a sample is taken from a sample container by a pipette 46 of the pipetting unit 41. The sample enters the pipette tip or needle of the pipette 46.

(65) Step E: The capture device is moved into the storage area 2. By the capture device 34, a sample carrier 6 is removed from the sample carrier receiving device 21, and 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 conveyed by the capture device 34 to the wetting device 31. In the process, it moves from the storage area 2 into the working area 3.

(66) Step F: The sample carrier 6 is conveyed by the capture device to a second identification device 50 (for example, a barcode reader), and identified there. The purpose of this is to allow a later assignment of the sample to the sample carrier 6.

(67) Step G: The first and second auxiliary materials are mixed to form an auxiliary material mixture. The auxiliary material mixture is the buffer. For the preparation of the auxiliary material mixture, defined amounts of the first and second auxiliary materials from the first and second auxiliary material receiving device 22, 23 are conveyed via the conveyor 33 to the wetting device 31 and mixed there to form a buffer. The defined amounts of the first and second auxiliary materials can correspond to a prespecified mixing ratio. It should be noted that step G is only required if no prefabricated buffer is used. In this case, the buffer is conveyed from an auxiliary material receiving device in which it is stored and cooled, via the conveyor 33, to the wetting device 31, where it can be used directly. It should also be noted that alternatively, the auxiliary material mixture can be prepared and the samples can be wetted with drops of the auxiliary material mixture by the pipetting system. This eliminates the wetting device 31 and/or a mixing device.

(68) Step H: The buffer is applied to the sample carrier 6 via the nozzles 32. In this case, the sample carrier 6 is rinsed with the buffer, and particularly 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.

(69) Steps I and I2: According to the specifications of the user of the apparatus, the following parameters are now set: the discharge height, the angle of inclination of the wetted sample carrier 6 resting on the heating/cooling plate 54, and optionally the discharge speed. To set the discharge height, the capture device 34 and the pipetting unit 41with the pipetting head 42 which holds the pipette with the pipette tipare aligned relative to each other. In this case, a prespecified capture region of the wetted sample carrier 6 moves into the capture position, and the pipette tip moves into the discharge position. The discharge height ha is preferably set to a value in a range from 0.1 to 800 mm; the inclination angle (double arrow c) of the wetted sample carrier 6 is set to a value in a range from 0 to 90; and the discharge speed is set to a value in the range from 0 to 10 m/s. The sample carrier is still on the heating/cooling plate.

(70) Step J: A prespecified amount of the sample is discharged in one or more drops 8 from the discharge position to the prespecified capture region 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 burst sample 9 and is therefore hereinafter referred to as the sample carrier 6. The sample carrier is optionally in a cooled state during the dropwise deposition of the sample. It can either be actively cooled or positioned on a cold block, or covered with cold buffer. To cover the sample carrier with cold buffer, the sample carrier can be immersed, for example, in the buffer.

(71) Step K: The sample carrier 6 is conveyed by the capture device 34 into a drying area. There, the angle of inclination of the sample carrier 6 is set to an angle of inclination of 0, in which the surface of the sample carrier 6 carrying the burst sample 9 lies in a horizontal plane. Subsequently, the sample carrier 6 carrying the wetted and burst sample 9 is heated by the heating/cooling plate 54 to a prespecified temperature for a prespecified period of time in order to achieve evaporation of the buffer and thus immobilization of the burst sample on the sample carrier 6. The temperature can be 90 C., by way of example, and the duration can be approx. 3 to 4 minutes. Optionally, the drying can be supported by fans. It should be noted that the cooling/heating position of the sample carrier 6 or 6 can be realized as one position, or as separate positions. In the latter case, rather than one heating/cooling plate 54, there can be a cooling plate in a first position and a heating plate in a second position, or the opposite: a heating plate in a first position and a cooling plate in a second position.

(72) Step L: This step is optional. The sample carrier 6 with the immobilized, burst sample is now conveyed by the capture device 34 to the analysis device 52. The analysis device 52 can be, for example, a microscope. By the analysis device 52, an automatedfor example, microscopicanalysis of the immobilized burst sample 9 can be carried out. In this case, the sample carrier 6 can be in a parked position.

(73) Step M: The pipetting head 41 with the pipette 42 which carries the pipette tip of the second type, used for taking up and discharging the sample, is conveyed to the collecting container 49. There, the pipette tip including the non-discharged portion of the sample is discarded. If a needle has been inserted instead of a pipette tip, the needle is ejected. The needle can then be washed and subsequently sterilized. If the needle is not discarded, it can optionally be washed, sterilized and reused in the apparatus.

(74) Step N: The sample carrier 6 with the immobilized, burst sample is now moved back into the storage area 2 by the capture device 34. There, the sample carrier 6 can be stored in the sample carrier receiving device 21.

(75) It is understood that the foregoing description is that of the exemplary embodiments of the disclosure and that various changes and modifications may be made thereto without departing from the spirit and scope of the disclosure as defined in the appended claims.

LIST OF REFERENCE NUMERALS

(76) 1 housing 2 cooled area 3 working area 3a working area sub-region 4 climatic chamber 5 computer 5a data connection 6 sample carrier 7 sample feed 8 drop of the sample 9 burst drop of the sample 21 sample carrier receiving device 22 first auxiliary material receiving device 23 second auxiliary material receiving device 24 third auxiliary material receiving device 31 wetting device 32 nozzle 33 conveying means 34 capture device 35 holding unit 36 actuator 37 arm 38 runner 39 rail 40 collecting unit 41 pipetting unit 42 pipetting head 43 arm 44 runner 45 rail 46 pipette 47 pipette tip holding device 48 sample receiving device 49 collecting vessel 50 identifying device 51 drying device 52 analysis device 53 supply means 54 heating/cooling plate