METHOD FOR ADJUSTING A CELL CONCENTRATION AND/OR A PARTICLE CONCENTRATION IN A DISPENSING SYSTEM

Abstract

The invention relates to a method for setting a cell concentration and/or a particle concentration in a dispensing device by means of which a liquid sample can be discharged, wherein the cell concentration and/or the particle concentration is ascertained in one region of the dispensing device, and the cell concentration and/or particle concentration that has been ascertained is compared with a target value, and a force exerted on one cell and/or one particle is adjusted based on the result of the comparison.

Claims

1. A method for setting a cell concentration and/or a particle concentration in a dispensing device (1) by means of which a liquid sample (20) can be discharged, the method comprising: ascertaining the cell concentration and/or the particle concentration in the liquid sample; comparing the cell concentration and/or particle concentration that has been ascertained with a target value; and adjusting a force exerted on one cell and/or one particle based on a result of the comparison.

2. The method according to claim 1, wherein the force is caused by sound and the force is adjusted by adjusting an orientation and/or a frequency and/or an amplitude and/or a phase and/or a modulation of the sound.

3. (canceled)

4. (canceled)

5. The method according to claim 1, wherein a triggering of the dispensing apparatus (6) is changed in order to adjust the cell concentration and/or particle concentration, wherein the triggering is changed by changing a specific time of the triggering and/or a frequency of the triggering and/or a manner of the triggering and/or an amplitude of the triggering and/or a volume of the discharged liquid sample (20) of the triggering.

6. (canceled)

7. The method according to claim 1, wherein the force is adjusted such that the cell and/or particle is held substantially stationary, and the liquid sample (20) is discharged by of the dispensing device (1) if the ascertained cell concentration and/or particle concentration is less than the target value, wherein the liquid sample (20) discharged by the dispensing device has liquid (2) and no cells and/or particles.

8. (canceled)

8. (canceled)

9. The method according to claim 1, wherein the force is adjusted such that movement of the cell and/or particle in a deploying direction (R) is possible if the ascertained cell concentration and/or particle concentration is equal to the target value.

10. The method according to claim 1, wherein if the cell concentration and/or particle concentration is greater than the target value, the liquid sample (20) is discharged.

11. (canceled)

12. The method according to claim 10, wherein a. the force is alternatingly adjusted to keep the cell and/or the particle substantially stationary, and then to exert no force on the cell and/or one particle, or b. the force is adjusted to oppose a movement of the cell and/or the particle, but the cell and/or the particle can move in a deploying direction (R).

13. (canceled)

14. The method according to claim 10, wherein at least one dispensing operation is carried out in which the discharged liquid sample (20) has liquid (2) and at least one cell (4) and/or at least one particle.

15. The method according to claim 1, wherein the dispensing device (1) has a discharge channel (3) through which the liquid sample (20) is discharged from the dispensing device, and the cell concentration and/or particle concentration is ascertained from a determination of the number of cells (4) and/or particles in at least a portion of the discharge channel (3) based on an image of the at least a portion of the discharge channel.

16. (canceled)

17. (canceled)

18. (canceled)

19. The method according to claim 15, wherein the force exerted on the cell and/or particle is provided by subjecting the at least a portion of the discharge channel (3) to an acoustic field.

20. The method according to claim 19, wherein the dispensing device (1) has a fluid chamber (5) for accommodating the liquid sample (20), the fluid chamber (5) being fluidically connected to the discharge channel (3), and wherein a. the fluid chamber (5) and/or the at least one portion of the discharge channel (3) is subjected to acoustic fields that differ from one another in frequency and/or amplitude and/or phase and/or modulation and/or orientation and/or wherein b. the fluid chamber (5) and/or at least a portion of the discharge channel (3) is subjected to acoustic fields of different frequency and/or amplitude and/or phase and/or modulation and/or orientation if the cell concentration and/or the particle concentration is less than the target value.

21. (canceled)

22. (canceled)

23. (canceled)

24. (canceled)

25. (canceled)

26. A dispensing apparatus (6) comprising: a dispensing device (1) including a fluid chamber (5) into which a liquid sample (20) is introduced and a discharge channel (3) fluidically connected to the fluid chamber (5) through which the liquid sample (20) is discharged from the dispensing device (1), wherein the liquid sample (21) includes a liquid (2) and cells (4) and/or particles; a sound generator (7) arranged in association with the dispensing device (1), the sound generator being operable to generate an acoustic field; an actuating means (8) for actuating the dispensing device (1) to cause the dispensing device (1) to discharge a volume of the liquid sample (20) from the dispensing device (1); an evaluation device (12) configured to ascertain a cell concentration and/or particle concentration in the dispensing device (1); and a control apparatus (9) connected to the dispensing device (1), the sound generator (7), the actuating means (8), and the evaluation device (12); wherein the control apparatus (9) is configured to compare the ascertained cell concentration and/or particle concentration with a target value to provide a comparison result, and configured to adjust the cell concentration and/or particle concentration in the dispensing device (1) by adjusting a force exerted on a cell (4) and/or a particle in the liquid sample (20) as a function of the comparison result.

27. (canceled)

28. The dispensing apparatus (6) according to claim 26, wherein the control apparatus (9) is configured to adjust the force such that the cell and/or particle is held substantially stationary, and to control the actuating means (8) such that the liquid sample (20) is discharged via the dispensing device (1) if the ascertained cell concentration and/or particle concentration is less than the target value.

29. The dispensing apparatus (6) according to claim 28, wherein the control apparatus (9) is configured to control the actuating means (8) such that a dispensing operation for discharging a volume of the liquid sample (20) having a liquid (2) and no cells and/or particles is carried out multiple times in succession.

30. The dispensing apparatus (6) according to claim 26, wherein the control apparatus (9) is configured to cause no force to be exerted on the cell and/or particle, or to adjust the force such that movement of the cell and/or particle in a discharge direction is possible if the ascertained cell concentration and/or particle concentration is equal to the target value.

31. (canceled)

32. (canceled)

33. (canceled)

34. (canceled)

35. The dispensing apparatus (6) according to claim 26, further comprising an imaging device (11) arranged to generate an image of at least a portion of the discharge channel (3) of the dispensing device (1), wherein the evaluation device (12) is configured to ascertain the number of cells (4) and/or particles present in the at least one portion of the discharge channel (3) based on the generated image.

36. (canceled)

37. The dispensing apparatus (6) according to claim 26, wherein the discharge channel (3) has at least one section having a flow cross-section that varies along a deploying direction (R) of the liquid sample (20).

38. The dispensing apparatus (6) according to claim 26, further comprising a diversion device (18) for deflecting the discharged liquid sample (20), wherein the control apparatus (9) is configured to control the diversion device (18) depending on the ascertained cell concentration and/or particle concentration.

39. The dispensing apparatus (6) according to claim 26, further comprising a displacement device (13), by means of which the dispensing device (1) and/or a container (14) for receiving the liquid sample (20) and/or a reject container (15) for receiving the liquid sample (20) can be displaced, wherein the control apparatus (9) is configured to control the displacement device (13) depending on the ascertained cell concentration and/or particle concentration.

40. A non-transient computer readable storage medium comprising a computer program comprising instructions that, when the computer program is executed by a computer (12), cause the computer to carry out the method according to claim 1.

41. (canceled)

42. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWING VIEWS

[0075] The subject matter of the disclosure is shown schematically in the figures, wherein elements that are the same or have the same effect are mostly provided with the same reference symbols. In the figures:

[0076] FIG. 1 shows a dispensing apparatus according to the disclosure,

[0077] FIG. 2 shows a dispensing device of the dispensing apparatus in a state in which the dispensing device is not subjected to an acoustic field,

[0078] FIG. 3 shows the dispensing device in a state in which the dispensing device is subjected to an acoustic field,

[0079] FIG. 4 shows the dispensing device in a state in which the dispensing device is subjected to an acoustic field and a liquid sample is discharged from the dispensing device,

[0080] FIG. 5 shows the dispensing device in a state in which the cell concentration in a portion of the discharge channel is too high,

[0081] FIG. 6 shows the dispensing device in a state in which liquid samples are discharged from the dispensing device,

[0082] FIG. 7 shows the dispensing device in a state in which the cells are concentrated centrally in the discharge channel and the cell concentration is too low,

[0083] FIG. 8 shows the dispensing device from FIG. 7 in a state in which multiple liquid drops are discharged,

[0084] FIG. 9 shows the dispensing device in a state in which the cells are concentrated centrally in the discharge channel and liquid samples are discharged,

[0085] FIG. 10 shows the dispensing device in a state in which the cells are aligned centrally in the discharge channel and liquid samples are discharged.

DETAILED DESCRIPTION

[0086] FIG. 1 shows a dispensing apparatus 6 according to the disclosure, which has a dispensing device 1 for discharging a liquid sample 20 that may have liquid 2 and at least one cell 4 and/or at least one particle. In addition, the dispensing apparatus 6 has an optical detection device 10 for optically detecting at least a portion of a discharge channel 3 of the dispensing device 1. The dispensing device 1 may have a fluid chamber 5 that contains liquid 2 and cells 4 and/or particles. The liquid chamber 5 is fluidically connected to the discharge channel 3.

[0087] The optical detection device 10 has an imaging device 11 for generating an image of the at least one portion of the discharge channel 3 and additional elements that are not shown in the drawings. To generate an image, the at least one part of the discharge channel 3 is illuminated by means of an illumination light 17 and a detection light 16 emanating from the at least one part of the discharge channel 3 is detected by the optical detection device 10. The imaging device 11 generates an image of the at least one portion of the discharge channel 3 based on the detected detection light 16.

[0088] The optical detection device 10 is electrically connected to an evaluation device 12. The evaluation device 12 can determine the number of cells 4 and/or particles contained in the at least one part of the discharge channel 3 based on the generated image. Thus, the evaluation device 12 can determine the cell concentration and/or particle concentration in the at least one portion of the discharge channel 3.

[0089] The evaluation device 12 is electrically connected to a control device 9. The control apparatus 9 and evaluation device 12 can be part of a computer. Based on the evaluation result of the evaluation device 12, the control apparatus 9 controls the dispensing operation of the dispensing device 1. The control apparatus 9 is electrically connected to a displacement device 13. The displacement device 13 can move the dispensing device 1 and/or the container 14 and/or a reject container 15 in such a way that the liquid 2 can be dispensed into the desired storage location.

[0090] In addition, the control apparatus 9 can control a deflection and/or suction device 18 of the dispensing apparatus 6. In this case, the control apparatus 9 can control the diversion device and/or suction device 18 in such a way that the discharged liquid sample 20 is diverted and/or suctioned away if the liquid sample 20 has no cells 4 and/or no particles, or if the liquid sample 20 has multiple cells 3 and/or multiple particles. Based on the cell concentration and/or particle concentration determined by the evaluation device 12, the control apparatus 9 can predict whether the liquid drop or liquid drops to be discharged during the next dispensing operation will have no cells and/or particles, or will have one or more cells and/or particles. The control apparatus 9 can then control the displacement device 13 and/or the diversion device and/or suction device 18 as a function of the prediction.

[0091] FIG. 1 shows a state in which the dispensing device 1 has discharged a liquid sample 20, in particular a liquid drop, that has a single cell 4. The liquid 2 together with the cell 4 is fed into the container 14, which is for example part of a microtitre plate that is not shown in greater detail.

[0092] The dispensing apparatus 6 has an actuating means 8 that is pressed against a section of the dispensing device 1 to actuate the dispensing device 1. In this case, the liquid sample 20, in particular a liquid drop, is discharged if the actuating means 8 presses against the section of the dispensing device 1. The actuating means 8 and the optical detection device 10 are opposite each other with respect to the dispensing device 1. The dispensing device 1 consists at least partially of a transparent material, such that at least a portion of the discharge channel 3 can be detected by means of the optical detection device 10.

[0093] The dispensing apparatus 1 also has a sound generator 7 that emits an acoustic field. In this case, the sound generator 7 is positioned in such a way that at least a portion of the discharge channel 3, in particular the entire discharge channel 3, can be subjected to the acoustic field. In particular, the sound generator 7 can be in mechanical contact with the actuating means 8 and thus can transmit the sound particularly efficiently.

[0094] FIG. 2 shows the dispensing device 1 of the dispensing apparatus 6 in a state in which the dispensing device 1 is not subjected to an acoustic field. In particular, FIG. 2 shows an enlarged view of the dispensing device 1 from the front.

[0095] The discharge channel 3 is completely filled with the liquid sample 20. In this case, only that portion of the discharge channel 3 that has a discharge opening of the discharge channel 3, shown in dashed lines in FIG. 2, is observed via the optical detection device 10. Three cells 4 of the liquid sample 20 are arranged in the observed portion of the discharge channel 3. During a dispensing process, the liquid sample 20 is discharged along a deploying direction R. The discharge channel 3 has a nozzle-shaped end at the end thereof remote from that of the fluid chamber 5. In addition, the discharge channel 3 has a nozzle-shaped end at its end facing towards the fluid chamber 5.

[0096] The cells 4 arranged in the discharge channel 3 move towards the discharge opening of the discharge channel 3 due to weight, even if no liquid 2 is discharged from the dispensing device 1.

[0097] FIG. 3 shows the dispensing device 1 in a state in which the dispensing device 1 is subjected to an acoustic field. In particular, the dashed portion of the discharge channel 3 is subjected to the acoustic field. By subjecting the portion of the discharge channel 3 to the acoustic field, the result is achieved that a force opposite the deploying direction R acts on the cells 4. Thus, the cells 4 do not move, or move to a lesser extent, towards the nozzle-shaped end due to the weight force. In this case, the cells 4 can be slowed or held substantially stationary, in particular stationary, in the deploying direction due to the force exerted on them. The cells 4 are aligned by the acoustic field in a direction transverse, in particular perpendicular, to the deploying direction R.

[0098] Since the upper end of the dispensing device 1 and the lower end of the dispensing device 1 have substantially the same design, by superimposing respectively one or more sound waves emitted from the sound generator 7 and one or more sound waves reflected on one or more discharge channel walls 19, one or, in the multidimensional case, multiple stationary waves can be generated, and the result can thus be achieved that the cells 4 are held substantially stationary, in particular stationary. The stationary waves run parallel to the deploying direction R and run between the upper end of the dispensing device 1 that faces towards the fluid chamber 5 and the lower end of the dispensing device 1 that faces away from the fluid chamber 5.

[0099] In the following, a first mode of operation of the dispensing apparatus 6 according to the disclosure is described with reference to FIGS. 1 to 4.

[0100] The imaging device 11 generates an image of the discharge channel 3, in particular the portion of the discharge channel 3 shown in dashed lines in FIG. 2. Based on the generated image, the evaluation device 12 determines the number of cells 4 contained in the dashed portion of the discharge channel 3 and thus determines the cell concentration in the observed portion of the discharge channel 3.

[0101] The control apparatus 9 checks whether the cell concentration is less than a predetermined target value. In the case shown in FIGS. 1-4, it is assumed that the cell concentration in the observed portion of the discharge channel 3 is too low, namely less than the target value.

[0102] In the next step, the sound generator 7 is activated and the discharge channel 3, in particular the portion of the discharge channel 3 shown in dashed lines, is subjected to an acoustic field generated by the sound generator 7. As described above, the acoustic field is configured in such a way that a force is exerted on the cells 4 to prevent the cells 4 from moving towards the nozzle-shaped end of the discharge channel 3 that faces away from the fluid chamber 5.

[0103] In addition, one dispensing operation is carried out, or multiple dispensing operations are carried out. In particular, multiple liquid drops are discharged, as shown in FIG. 4. In addition, liquid drops are discharged that exclusively contain liquid 2 and thus do not contain any cells and/or particles. Since a force directed in the direction opposite the deploying direction R acts on the cells 4 that are arranged in the portion of the discharge channel 3, the cells 4 remain in their position during the dispensing operation or operations. The liquid discharge causes a portion of the fluid present in the fluid chamber 5 to flow into the discharge channel 3 via the upper end of the dispensing device 1. Since the liquid 2 present in the fluid chamber 5 contains cells 4, new cells 4 thus enter the observed portion of the discharge channel 3. Specifically, in the embodiment shown in FIG. 4, two new cells 4 enter the observed portion of the discharge channel 3, such that the cell concentration in the observed portion of the discharge channel 4 has been increased. Moreover, four additional cells 4 from the fluid chamber 5 reach the non-observed portion of the discharge channel 3. Subsequently, as described above, the cell concentration is again determined and if necessary the above-described steps are repeated until the cell concentration has reached the target value.

[0104] Below, a second mode of operation of the dispensing apparatus 6 according to the disclosure is described with reference to FIGS. 1, 5 and 6. As in the first mode of operation, the cell concentration is first ascertained in the portion of the discharge channel 3 shown in dashed lines in FIG. 5. The control apparatus 9 then ascertains that the cell concentration is greater than the target value.

[0105] In the next step, the sound generator 7 is switched off and the actuating means 8 is actuated such that a dispensing operation is carried out. After the liquid sample 20 has been discharged, the sound generator 7 is switched back on, and the observed portion of the discharge channel 3 is subjected to the acoustic field. Switching the sound generator on and off before the liquid sample 20 is discharged is repeated multiple times in a predetermined period of time and/or for a predetermined number of dispensing operations.

[0106] Alternatively, the sound generator 7 in the switched-on state is set in such a way that the force exerted on the cells causes the cells to be held substantially stationary, in particular stationary. Subsequently, the sound generator 7 is not switched off, but is shifted to a second state in which the force acting on the cells is less than the flow force acting on the cells during the dispensing operation. In addition, a dispensing operation is carried out if the sound generator 7 is in the second state. The force acting on the cells in the second state of the sound generator may be greater than the weight force acting on the cells, but less than the flow force that acts on the cells during the dispensing operation. Consequently, the movement of the cells towards the discharge opening is slowed.

[0107] Regardless of which of the above-mentioned methods is used to operate the sound generator 7, a plurality of discharged liquid samples 20 have only a single cell 4, as is apparent in FIG. 6. After the predetermined time period and/or the predetermined number of dispensing operations, the cell concentration and/or particle concentration is checked again. The sound generator is switched on again if the cell concentration is greater than the target value. The above-mentioned steps are repeated multiple times until the cell concentration is equal to the target value or less than the target value.

[0108] Since, in the state shown in FIG. 6, some cells 4 have been discharged compared to the state shown in FIG. 5 and thus are no longer arranged in the observed portion of the discharge channel 3, the cell concentration in the observed portion of the discharge channel 3 is lower than prior to the dispensing operations.

[0109] FIGS. 7 to 10 show a third mode of operation of the dispensing apparatus according to the disclosure. The modes of operation described in FIGS. 7 to 10 share the fact that the cells 4 that are present in the discharge channel 3 are concentrated or aligned in the discharge channel 3 using an acoustic field. In addition, and in particular subsequently, the cell and/or particle concentration is adjusted. The cell and/or particle concentration can be adjusted by the corresponding region of the discharge channel 3 being subjected to a different acoustic field. Alternatively, the cell concentration and particle concentration can be adjusted by changing the setting of the acoustic field that causes the cells to be concentrated or aligned, such as for example the frequency, amplitude, etc., of the acoustic field. In this case, it is not necessary that multiple acoustic fields are applied to the discharge channel, but instead, with the same acoustic field, concentration or alignment can be effected and the cell concentration and particle concentration can be adjusted.

[0110] FIGS. 7 and 8 show a state in which the cells are concentrated in a central portion of the discharge channel 3. The cells are concentrated in the central region of the discharge channel 3 as a result of the discharge channel 3 being subjected to an acoustic field. The acoustic field is adjusted in such a way that the cells are respectively subjected to a force that causes them to be concentrated in the central region of the discharge channel 3 shown in FIG. 7.

[0111] In this case, the cell concentration in the observed region of the discharge channel 3, which is not shown in FIGS. 7 and 8, is less than the target value. Therefore, the acoustic field is adjusted in such a way that the cells 4 arranged in the observed region of the discharge channel 3 are not moved in the deploying direction R. In particular, the acoustic field is adjusted in such a way that the cells 4 arranged in the discharge channel 3 do not move even after multiple of the liquid drops shown in FIG. 8 have been discharged, and are thus substantially stationary.

[0112] In contrast, as is apparent from FIG. 8, as a result of the dispensing operations, some of the cells 4 arranged outside the observed region of the discharge channel 3 in FIG. 7, which for example were arranged in the fluid chamber 5 before the dispensing operation, enter the observed region, such that the cell concentration in the observed region of the discharge channel 3 increases.

[0113] In the embodiment shown in FIG. 9, the cells 4 are concentrated in the centre of the discharge channel 3 owing to the applied acoustic field. Although not shown in FIG. 9, in the embodiment the cell concentration in the observed portion, not shown, of the discharge channel 3 was too high, namely above the target value. Thus, analogously to the mode of operation described with reference to the second mode of operation, the sound generator is alternatingly switched on and off or adjusted (for example by reducing the force on the cells) so that only some of the cells are supplied to the observed region, such that the cell concentration in the observed region is reduced and the probability is increased that the discharged liquid samples 20 each respectively have a single cell 4.

[0114] In the embodiment shown in FIG. 10, the cells 4 are aligned centrally owing to the adjacent acoustic field. The cell concentration in the observed portion, not shown, of the discharge channel 3 is then equal to the target value. In this case, the acoustic field does not need to be further modified to yield a change in cell concentration in the observed portion of the discharge channel 3.

[0115] In each of the above-described modes of operation, an image is generated of the discharge channel 3, in particular of the portion of the discharge channel 3, and the number of cells 4 and/or particles arranged in the discharge channel 3 is determined. Depending on the number of cells 4 present in the discharge channel 3, the observed portion of the discharge channel 3 is or is not subjected to the acoustic field.

LIST OF REFERENCE SIGNS

[0116] 1 Dispensing device

[0117] 2 Liquid

[0118] 3 Discharge channel

[0119] 4 Cell

[0120] 5 Fluid chamber

[0121] 6 Dispensing apparatus

[0122] 7 Sound generator

[0123] 8 Actuating means

[0124] 9 Control apparatus

[0125] 10 Detection device

[0126] 11 Imaging device

[0127] 12 Evaluation apparatus

[0128] 13 Displacement device

[0129] 14 Container

[0130] 15 Reject container

[0131] 16 Detection light

[0132] 17 Illumination light

[0133] 18 Deflection and/or suction device

[0134] 19 Discharge channel wall

[0135] 20 Liquid sample

[0136] R Deploying direction