Single cell suction device and method

Abstract

The invention discloses a single cell suction device and method, and relates to the technical field of single cell separation.The single cell suction device comprises a multi-axis adjusting platform, a suction mechanism, a sample moving platform, a sample holder, a focusing module and an imaging system.The output end of the multi-axis adjusting platform is connected with the suction mechanism, and the output end of the sample moving platform is connected with the sample holder; the suction mechanism is located above the sample holder, the sample holder is used for placing a single cell sample, the focusing module is located below the sample moving platform and can focus the sample holder and the tail end of the suction mechanism, and the imaging system is located below the sample moving platform and can focus the sample holder and the tail end of the suction mechanism. The multi-axis adjusting platform is used for moving the tail end of the suction mechanism to the center of the imaging system, the sample moving platform is used for driving the sample rack to move and enabling the single-cell sample on the sample rack to be located at the center of the imaging system, and the suction mechanism is used for sucking the single-cell sample on the sample rack. According to the invention, automatic needle alignment and automatic stable suction and release of target cells can be realized, and the operation difficulty is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of single cell separation, and in particular to a single cell aspiration device and method. Background Art

[0002] At present, the commonly used methods for separating single cells include infinite dilution method, micromanipulation method and laser microdissection technology. Among them, micromanipulation method and laser microdissection technology can directionally separate target single cells. For laser microdissection technology, there are defects such as high cost and easy damage to cells, so the most common single cell picking method is micromanipulation method. However, the micromanipulation method often uses a mouth aspirator to manually suck up single cells, which has the disadvantages of poor stability and high difficulty in operation. In addition, the micromanipulation table requires manual needle alignment, which has high technical requirements for the operator. Summary of the invention

[0003] The purpose of the present invention is to provide a single cell aspiration device and method to solve the problems existing in the above-mentioned prior art, realize automatic needle alignment and automatic stable aspiration and release of target cells, and reduce the difficulty of operation.

[0004] To achieve the above object, the present invention provides the following solutions:

[0005] The present invention provides a single-cell suction device, comprising a multi-axis adjustment platform, a suction mechanism, a sample moving platform, a sample rack, a focusing module and an imaging system, wherein the output end of the multi-axis adjustment platform is connected to the suction mechanism, the output end of the sample moving platform is connected to the sample rack, the suction mechanism is located above the sample rack, and the sample rack is used to place single-cell samples, the focusing module is located below the sample moving platform, and the focusing module can focus on the sample rack and the end of the suction mechanism, the imaging system is located below the sample moving platform, and the multi-axis adjustment platform is used to move the end of the suction mechanism to the center of the imaging system, the sample moving platform is used to drive the sample rack to move, and make the single-cell sample on the sample rack located at the center of the imaging system, and the suction mechanism is used to suck the single-cell sample on the sample rack.

[0006] Preferably, the multi-axis adjustment platform is a three-axis adjustment platform, and the three-axis adjustment platform can drive the suction mechanism to achieve reciprocating movement in directions A, B and C.

[0007] Preferably, the suction mechanism includes a negative pressure element and a suction head, wherein the suction head is installed at the output end of the multi-axis adjustment platform, the negative pressure element is connected to one end of the suction head, and the negative pressure element is used to provide negative pressure suction for the suction head, and the other end of the suction head is used to suck the single cell sample on the sample rack.

[0008] Preferably, the negative pressure element is an electric microinjection pump, and the suction head is a capillary suction needle.

[0009] Preferably, the electric microinjection pump is connected to the capillary sampling needle via a pipeline.

[0010] Preferably, the sample moving platform is a two-axis moving platform, and the two-axis moving platform can drive the sample holder to move back and forth along the X direction and the Y direction.

[0011] Preferably, a plurality of sample holes are provided at the upper end of the sample rack, and each of the sample holes is used to place a single cell sample.

[0012] Preferably, the focusing module is a Z-axis objective lens focusing module, and the Z-axis objective lens focusing module is used to focus on the Z plane.

[0013] Preferably, the imaging system is an inverted microscopic imaging system.

[0014] The present invention also provides a single cell aspiration method, using the single cell aspiration device described in any one of the above technical solutions, comprising the following steps:

[0015] S1. Place a sample holder without a single cell sample on the sample moving platform, use the focusing module to focus and record the plane where the upper surface of the sample holder is located;

[0016] S2. Move the capillary sampling needle in the aspiration mechanism to the center of the field of view of the imaging system through the multi-axis adjustment platform, and at this time, the lower end of the capillary sampling needle is higher than the upper surface of the sample rack, or the lower end of the capillary sampling needle is flush with the upper surface of the sample rack, record the coordinates of the capillary sampling needle at this time, and then lift the capillary sampling needle;

[0017] S3. Remove the sample rack without single-cell samples, and place the sample rack with single-cell samples on the sample moving platform, use the focusing module to focus on the plane where the upper surface of the target single-cell sample is located, and adjust the sample moving platform to move the target single-cell sample to the center of the field of view of the imaging system;

[0018] S4. Use the multi-axis adjustment platform to lower the capillary sampling needle to the upper surface of the sample holder;

[0019] S5. Adjust the electric micro-injection pump in the suction mechanism so that the electric micro-injection pump controls the capillary suction needle to suck the single-cell sample.

[0020] Compared with the prior art, the present invention has achieved the following technical effects:

[0021] The single-cell suction device and method provided by the present invention are characterized in that the output end of the multi-axis adjustment platform is connected to the suction mechanism, the output end of the sample moving platform is connected to the sample rack, the suction mechanism is located above the sample rack, and the sample rack is used to place the single-cell sample, the focusing module is located below the sample moving platform, and the focusing module can focus on the sample rack and the end of the suction mechanism, and then through the arrangement of the multi-axis adjustment platform and the sample moving platform, the end of the suction mechanism is electrically controlled to align with the single-cell sample, and the relative height of the focusing plane at the end of the suction mechanism and the focusing plane of the single-cell sample is controlled respectively, so that the focusing plane at the end of the suction mechanism is higher than the focusing plane of the single-cell sample, or the focusing plane at the end of the suction mechanism is aligned with the single-cell sample. The focal plane is flush with the focal plane of the single-cell sample to avoid needle collision, which affects the service life. The imaging system is located below the sample moving platform, and the multi-axis adjustment platform is used to move the end of the suction mechanism to the center of the imaging system. The sample moving platform is used to drive the sample rack to move and make the single-cell sample on the sample rack located in the center of the imaging system, and the suction mechanism is used to suck the single-cell sample on the sample rack. The relative position of the end of the suction mechanism and the single-cell sample in the three-dimensional space is adjusted through automatic control, thereby realizing automatic suction and collection of single-cell samples. Compared with traditional manual needle alignment, it not only reduces the difficulty of operation, but also reduces operation errors. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1 is a schematic diagram of the structure of the single cell aspiration device in Example 1;

[0024] Figure 2 It is a schematic diagram of the positions of various planes in the specific application embodiment 1;

[0025] Figure 3 It is a schematic diagram of the positions of various planes in the specific application embodiment 2;

[0026] Figure 4 It is a schematic diagram of the positions of various planes in the specific application embodiment 3;

[0027] In the figure: 1-multi-axis adjustment platform, 2-capillary sampling needle, 3-sample moving platform, 4-sample holder, 5-focusing module, 6-imaging system, 7-electric micro-injection pump. DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] The purpose of the present invention is to provide a single cell aspiration device and method to solve the problems existing in the prior art, realize automatic needle alignment and automatic stable aspiration and release of target cells, and reduce the difficulty of operation.

[0030] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0031] Embodiment 1

[0032] like Figure 1 As shown, this embodiment provides a single-cell suction device, including a multi-axis adjustment platform 1, a suction mechanism, a sample moving platform 3, a sample rack 4, a focusing module 5 and an imaging system 6, wherein the output end of the multi-axis adjustment platform 1 is connected to the suction mechanism, the output end of the sample moving platform 3 is connected to the sample rack 4, the suction mechanism is located above the sample rack 4, and the sample rack 4 is used to place the single-cell sample, the focusing module 5 is located below the sample moving platform 3, and the focusing module 5 can focus on the sample rack 4 and the end of the suction mechanism, and then through the arrangement of the multi-axis adjustment platform 1 and the sample moving platform 3, the end of the suction mechanism is electrically controlled to align with the single-cell sample, and the relative heights of the focusing plane at the end of the suction mechanism and the focusing plane of the single-cell sample are controlled respectively, so that the end of the suction mechanism is focused The plane is higher than the focusing plane of the single-cell sample, or the focusing plane of the end of the suction mechanism is flush with the focusing plane of the single-cell sample to avoid the needle collision phenomenon and affect the service life. The imaging system 6 is located below the sample moving platform 3, and the multi-axis adjustment platform 1 is used to move the end of the suction mechanism to the center of the imaging system 6. The sample moving platform 3 is used to drive the sample rack 4 to move and make the single-cell sample on the sample rack 4 located at the center of the imaging system 6, and the suction mechanism is used to absorb the single-cell sample on the sample rack 4. The relative position of the end of the suction mechanism and the single-cell sample in the three-dimensional space is adjusted through automatic control, thereby realizing automatic suction and collection of single-cell samples. Compared with the traditional manual needle alignment, it not only reduces the difficulty of operation, but also reduces the operation error.

[0033] Specifically, the multi-axis adjustment platform 1 is a three-axis adjustment platform, and the three-axis adjustment platform can drive the suction mechanism to achieve reciprocating movement in directions A, B and C, wherein direction A is the left and right direction, direction B is the front and back direction, and direction C is the up and down direction. The three-axis adjustment platform is preferably fine-tuned by controlling the screw mechanism through a precision stepping motor.

[0034] The suction mechanism includes a negative pressure element and a suction head. The suction head is installed at the output end of the multi-axis adjustment platform 1. The negative pressure element is connected to one end of the suction head, and the negative pressure element is used to provide negative pressure suction for the suction head. The other end of the suction head is used to absorb the single cell sample on the sample rack 4 to achieve non-destructive suction of the single cell sample.

[0035] The negative pressure element is preferably an electric micro-injection pump 7, and the suction head is preferably a capillary sample suction needle 2. The electric micro-injection pump 7 is connected to the capillary sample suction needle 2 through a pipeline. The electric micro-injection pump 7 is used as the suction power source for the single-cell sample. The adjustable suction interval is small. The software intelligently judges the suction of the single-cell sample as a feedback mechanism to change the pulse frequency of the electric micro-injection pump 7, so that the suction can be adjusted to make the sample suction more stable. During the operation, it is necessary to keep the needle tip of the capillary sample suction needle 2, the target single-cell sample and the center of the field of view on the same vertical line to achieve the smooth absorption of the target single-cell sample through the capillary sample suction needle 2.

[0036] The sample moving platform 3 is a two-axis moving platform, which can drive the sample holder 4 to move back and forth along the X direction and the Y direction, and the sample holder 4 cannot move in the Z direction, that is, the upper surface of the sample holder 4 is a fixed plane. The X direction is the left-right direction, and the Y direction is the front-back direction.

[0037] The upper end of the sample rack 4 is provided with a plurality of sample holes, each of which is used to place a single cell sample.

[0038] The focusing module 5 is a Z-axis objective lens focusing module, and the focusing module 5 includes an electric objective lens conversion component. The Z-axis objective lens focusing module is used to focus on the Z plane to achieve the upper and lower focusing of the sample holder 4 and the capillary sampling needle 2.

[0039] The imaging system 6 is an inverted microscopic imaging system.

[0040] Embodiment 2

[0041] This embodiment provides a single cell aspiration method, using the single cell aspiration device in Embodiment 1, comprising the following steps:

[0042] S1. Place a sample holder 4 without a single cell sample on the sample moving platform 3, and use the focusing module 5 to focus and record the plane where the upper surface of the sample holder 4 is located;

[0043] S2. Move the capillary sample aspiration needle 2 in the aspiration mechanism to the center of the field of view of the imaging system 6 through the multi-axis adjustment platform 1, and at this time, the lower end of the capillary sample aspiration needle 2 is higher than the upper surface of the sample holder 4, or the lower end of the capillary sample aspiration needle 2 is flush with the upper surface of the sample holder 4, record the coordinates of the capillary sample aspiration needle 2 at this time, and then lift the capillary sample aspiration needle 2;

[0044] S3. Remove the sample rack 4 without the single-cell sample, and place the sample rack 4 with the single-cell sample on the sample moving platform 3, use the focusing module 5 to focus on the plane where the upper surface of the target single-cell sample is located, and adjust the sample moving platform 3 to move the target single-cell sample to the center of the field of view of the imaging system 6;

[0045] S4. Use the multi-axis adjustment platform 1 to lower the capillary sampling needle 2 to the upper surface of the sample holder 4;

[0046] S5. Adjust the electric micro-injection pump 7 in the suction mechanism so that the electric micro-injection pump 7 controls the capillary sampling needle 2 to suck the single cell sample.

[0047] In specific applications, specific application examples of the single cell aspiration device in Embodiment 1 include but are not limited to the following:

[0048] Specific application example 1

[0049] like Figure 2 As shown, this specific application embodiment includes the following steps:

[0050] (1) Place a sample holder 4 without a single cell sample on the sample moving platform 3, and control the focusing module 5 to focus on the plane Z on the sample holder 4. 1 , then lift the focus module 5 to Z 2 , at this time Z 2 Plane above Z 1 flat;

[0051] (2) Control the multi-axis adjustment platform 1 to move the capillary sample needle 2 to the center of the imaging field of view, focus on the tip of the capillary sample needle 2, and record the position coordinates of the tip of the capillary sample needle 2 at this time (A 1 , B 1 , C 1 ), at this time C 1 Plane and Z 2 The planes are on the same horizontal plane. Keep the multi-axis adjustment platform 1 stationary in the A-axis and B-axis directions, move in the C-axis direction, and lift the capillary sampling needle 2 to the origin position C. 0 ;

[0052] (3) Remove the blank sample holder 4 and place the sample holder 4 with the single cell sample on the sample moving platform 3, and control the focusing module 5 to focus on the plane Z of the target single cell sample again.1 , control the movement of the sample moving platform 3 to move the target single cell sample to the center of the imaging field of view, so that the tip of the capillary aspiration needle 2, the target single cell sample and the center of the imaging field of view are on the same vertical line;

[0053] (4) Control the multi-axis adjustment platform 1 to move in the C direction and lower the capillary sampling needle 2 to the C direction. 1 , at this time C 1 Plane above Z 1 The plane can finely adjust the height of the tip of the capillary sampling needle 2 so that the tip fits the target single cell sample;

[0054] (5) Adjust the parameters of the electric microinjection pump 7 to perform aspiration.

[0055] like Figure 3 As shown, the specific application example 2

[0056] This specific application embodiment includes the following steps:

[0057] (1) Place a sample holder 4 without a single cell sample on the sample moving platform 3, and control the focusing module 5 to focus on the plane Z on the sample holder 4. 1 , and then remove the sample rack 4 that does not contain the single cell sample;

[0058] (2) Control the multi-axis adjustment platform 1 to move the capillary sampling needle 2 to the center of the imaging field of view, focus the tip of the capillary sampling needle 2, and record the position coordinates of the tip at this time (A 1 , B 1 , C 1 ), at this time C 1 Plane and Z 1 The planes are on the same horizontal plane. Keep the multi-axis adjustment platform 1 stationary in the A-axis and B-axis directions, move in the C-axis direction, and lift the capillary sampling needle 2 to the origin position C. 0 ;

[0059] (3) Place the sample holder 4 with the single cell sample on the sample moving platform 3, and control the focusing module 5 to focus on the plane Z of the target single cell sample again. 1 , control the movement of the sample moving platform 3 to move the target single cell sample to the center of the imaging field of view, so that the tip of the capillary aspiration needle 2, the target single cell sample and the center of the imaging field of view are on the same vertical line;

[0060] (4) Set up a C 2 plane, so that C 2 Plane above C 1 plane (difference is between 100 steps and 2000 steps), control the multi-axis adjustment platform 1 to move in the C-axis direction, and lower the capillary sampling needle 2 to the C 2 high;

[0061] (5) Set the parameters of the electric microinjection pump 7 to control the multi-axis adjustment platform 1 to move in the C-axis direction and lower the capillary sample aspiration needle 2 to the C-axis direction. 1 Absorb at high altitude.

[0062] Specific application example three

[0063] like Figure 4 As shown, this specific application embodiment includes the following steps:

[0064] (1) Place a sample holder 4 without a single cell sample on the sample moving platform 3, and control the focusing module 5 to focus on the plane Z on the sample holder 4. 1 , record Z 1 , and control the focus module 5 to lower the sample moving platform 3 as a whole to Z 2 ;

[0065] (2) Control the multi-axis adjustment platform 1 to move the capillary sample needle 2 to the center of the imaging field of view, focus on the tip of the capillary sample needle 2, and record the position coordinates of the tip of the capillary sample needle 2 at this time (A 1 , B 1 , C 1 ), at this time C 1 Plane and Z 1 The planes are on the same horizontal plane. Keep the multi-axis adjustment platform 1 stationary in the A-axis and B-axis directions, move in the C-axis direction, and lift the capillary sampling needle 2 to the origin position C. 0 ;

[0066] (3) Remove the blank sample holder 4 and place the sample holder 4 with the single cell sample on the sample moving platform 3, and control the focusing module 5 to lift the sample moving platform 3 to the plane Z. 1 , at this time Z 1 The plane is the focus plane of the target single cell sample, and the sample moving platform 3 is adjusted to move the target single cell sample to the center of the field of view;

[0067] (4) Set up a C 2 plane, so that C 2 Plane above C 1 plane (difference is between 100 steps and 2000 steps), control the multi-axis adjustment platform 1 to move in the C-axis direction, and lower the capillary sampling needle 2 to the C 2 high;

[0068] (5) Set the parameters of the electric microinjection pump 7 to control the multi-axis adjustment platform 1 to move in the C-axis direction and lower the capillary sample aspiration needle 2 to the C-axis direction. 1 Absorb at high altitude.

[0069] The present invention uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only used to help understand the method and core ideas of the present invention. At the same time, for those skilled in the art, according to the ideas of the present invention, there will be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A single cell aspiration device, characterized in that: The invention comprises a multi-axis adjustment platform, a suction mechanism, a sample moving platform, a sample rack, a focusing module and an imaging system, wherein the output end of the multi-axis adjustment platform is connected to the suction mechanism, the output end of the sample moving platform is connected to the sample rack, the suction mechanism is located above the sample rack, and the sample rack is used to place single-cell samples, the focusing module is located below the sample moving platform, and the focusing module can focus on the sample rack and the end of the suction mechanism, the imaging system is located below the sample moving platform, and the multi-axis adjustment platform is used to move the end of the suction mechanism to the center of the imaging system, the sample moving platform is used to drive the sample rack to move, and make the single-cell sample on the sample rack located at the center of the imaging system, and the suction mechanism is used to suck the single-cell sample on the sample rack.

2. The single cell aspiration device according to claim 1, characterized in that: The multi-axis adjustment platform is a three-axis adjustment platform, and the three-axis adjustment platform can drive the suction mechanism to achieve reciprocating movement in directions A, B and C.

3. The single cell aspiration device according to claim 1, characterized in that: The suction mechanism includes a negative pressure element and a suction head, wherein the suction head is installed at the output end of the multi-axis adjustment platform, the negative pressure element is connected to one end of the suction head, and the negative pressure element is used to provide negative pressure suction for the suction head, and the other end of the suction head is used to suck the single cell sample on the sample rack.

4. The single cell aspiration device according to claim 3, characterized in that: The negative pressure element is an electric micro-injection pump, and the suction head is a capillary sample suction needle.

5. The single cell aspiration device according to claim 4, characterized in that: The electric micro-injection pump is connected to the capillary sampling needle through a pipeline.

6. The single cell aspiration device according to claim 1, characterized in that: The sample moving platform is a two-axis moving platform, and the two-axis moving platform can drive the sample holder to move back and forth along the X direction and the Y direction.

7. The single cell aspiration device according to claim 1, characterized in that: A plurality of sample holes are arranged at the upper end of the sample rack, and each of the sample holes is used for placing a single cell sample.

8. The single cell aspiration device according to claim 1, characterized in that: The focusing module is a Z-axis objective lens focusing module, and the Z-axis objective lens focusing module is used to focus on the Z plane.

9. The single cell aspiration device according to claim 1, characterized in that: The imaging system is an inverted microscopic imaging system.

10. A single cell aspiration method, characterized in that: Using the single cell aspiration device according to any one of claims 1 to 9 comprises the following steps: S1. Place a sample holder without a single cell sample on the sample moving platform, use the focusing module to focus and record the plane where the upper surface of the sample holder is located; S2. Move the capillary sampling needle in the aspiration mechanism to the center of the field of view of the imaging system through the multi-axis adjustment platform, and at this time, the lower end of the capillary sampling needle is higher than the upper surface of the sample rack, or the lower end of the capillary sampling needle is flush with the upper surface of the sample rack, record the coordinates of the capillary sampling needle at this time, and then lift the capillary sampling needle; S3. Remove the sample rack without single-cell samples, and place the sample rack with single-cell samples on the sample moving platform, use the focusing module to focus on the plane where the upper surface of the target single-cell sample is located, and adjust the sample moving platform to move the target single-cell sample to the center of the field of view of the imaging system; S4. Use the multi-axis adjustment platform to lower the capillary sampling needle to the upper surface of the sample holder; S5. Adjust the electric micro-injection pump in the suction mechanism so that the electric micro-injection pump controls the capillary suction needle to suck the single-cell sample.

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