Single cell analysis system and method based on mass spectrometry technology

Abstract

The invention provides a single cell analysis system and method based on a mass spectrometry technology, the single cell analysis system comprises a support, an extraction platform and a mass spectrometry analyzer, the support is provided with a plurality of electrospray modules; the electrospray module comprises a base, a conductive probe and a capillary tube, and the transfer position is used for temporarily storing the electrospray module; the three-axis electric claw is used for grabbing the electrospray module on the bracket and transferring the electrospray module to the transfer position; the grabbing unit is used for grabbing the electrospray module at the transfer position, moving the electrospray module to an extraction platform for cell extraction, and then placing the electrospray module subjected to cell extraction on the ionization unit; the ionization unit is used for bearing the electrospray module and adjusting the position of the electrospray module, and an outlet of the capillary corresponds to a sample injection conical opening of the mass spectrum analyzer. The device has the advantages of automation, simple structure and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to cell analysis, in particular to a single cell analysis system and method based on mass spectrometry technology. BACKGROUND

[0002] At present, mass spectrometry has extremely wide applications in the fields of environmental detection, clinical analysis, organic synthesis, drug research and development, protein and metabolomics. The current electrospray device of mass spectrometry usually has the following technical problems: 1. The manual loading structure of the electrospray module cannot meet the continuous automatic testing.

[0003] 2. The position of the electrospray needle tip and the mass spectrometry cone cannot be accurately adjusted automatically, which reduces the quality of the electrospray sprayed into the cone.

[0004] 3. In the grabbing of the electrospray module, an open-close type electric claw is usually used, which has low position accuracy and cannot realize automatic centering.

[0005] 4. The electrification effect of the electrospray module is not good, and it is difficult to form a stable ionization state. SUMMARY

[0006] In order to solve the above problems in the prior art, the present application provides a single cell analysis system based on mass spectrometry technology.

[0007] The purpose of the present application is achieved by the following technical solutions: The single cell analysis system based on mass spectrometry technology comprises a support, an extraction platform and a mass spectrometer, a plurality of electrospray modules are arranged on the support, and the extraction platform and the mass spectrometer are arranged on a base; the electrospray module comprises a base, a conductive probe and a capillary tube, the capillary tube is inserted into the base, and the conductive probe is inserted into the capillary tube; the single cell analysis system further comprises: a transfer position, which is used for temporarily storing the electrospray module; a three-axis electric claw, which is used for grabbing the electrospray module on the support and transferring it to the transfer position; a grabbing unit, which is used for grabbing the electrospray module in the transfer position and moving to the extraction platform for cell extraction, and then placing the electrospray module after cell extraction on an ionization unit; an ionization unit, which is used for carrying the electrospray module and adjusting the position thereof, so that the outlet of the capillary tube corresponds to the sampling cone of the mass spectrometer.

[0008] Another purpose of the present application is to provide a single cell mass spectrometry analysis method, and the purpose of the present application is achieved by the following technical solutions: According to the single cell analysis method of the single cell analysis system of the application, the single cell analysis method comprises the following steps: (A1) The three-axis electric claw grabs the electrospray module on the support and places it in the transfer position; (A2) The grabbing unit grabs the electrospray module at the transfer position and moves to the extraction platform; (A3) The capillary of the electrospray module is used to complete the visualization of the selected cell on the extraction platform, and the capillary sucks the extraction material; (A4) The grabbing unit continues to move the electrospray module and places the electrospray module on the ionization unit; (A5) The ionization unit adjusts the position of the electrospray module through translation and rotation, so that the outlet of the capillary corresponds to the sample inlet orifice of the mass spectrometer; (A6) The electrically conductive probe in the capillary discharges, the extraction material is ionized, and the ions enter the sample inlet orifice.

[0009] Compared with the prior art, the application has the beneficial effects that: 1. Automation, high efficiency of single cell analysis; The electrospray module starts from the support, and sequentially experiences transfer, cell extraction, ionization and discarding, and these operations are automatically completed, thereby significantly improving the efficiency of single cell analysis; With the provided support, 20 or more electrospray modules can be installed on each support, meeting the requirements of rapid and large quantity loading; The transverse opening and longitudinal channel design of the bearing position enables the electrospray module to be quickly and accurately placed on the support, improving the overall analysis efficiency; 2. Good working stability; In the electrospray module, the protection part design effectively avoids damage to the capillary needle tip due to collision; the magnetic attraction structure and the ball head plunger structure ensure the positioning accuracy of each electrospray module after loading, and can keep the support fixed under the normal running vibration of the single cell analysis system based on mass spectrometry technology; The combination of the carbon steel round tube in the electrospray module and the magnet on the fixed seat in the ionization unit is used to position the electrospray module, improving the positioning accuracy of the electrospray module; In the grabbing unit, the moving part is provided with a three-section bearing structure, so that the electrospray module easily enters and is carried by the smaller inner diameter section, and the base is in the larger inner diameter section, improving the positioning accuracy; The electrically conductive probe passes through the guide hole between the deformation and is tightly clamped, and is not easy to loosen and fall off; 3. Simple structure; In the ionization unit, the three-dimensional movement of the moving platform, including a plurality of guide rails, driving wheels and driven wheels, etc. are all conventional components; a plurality of rotating modules are also conventional components, which are combined together to play the roles of placing, electrically conducting, three-dimensionally moving to the cone and automatically unloading the electrospray module of the electrospray module. A plurality of angle macro lenses take pictures, algorithms are used to identify the images, and then the positions of X, Y and Z three axes are automatically adjusted to realize the accurate and stable positioning of the ion source needle tip to the mass spectrometry cone.

[0010] The disclosure of the present application will become more apparent with reference to the drawings. It is easy for those skilled in the art to understand that the drawings are only used to illustrate the technical solutions of the present application, and are not intended to limit the protection scope of the present application. In the drawings: Figure 1 is a structural schematic diagram of a single cell analysis system based on mass spectrometry technology according to an embodiment of the present application; Figure 2 is a structural schematic diagram of an extraction platform according to an embodiment of the present application; Figure 3 is a structural schematic diagram of a bracket according to an embodiment of the present application; Figure 4 is a sectional view schematic diagram of an electrospray module according to an embodiment of the present application; Figure 5 is a schematic diagram of the bracket when the electrospray module is installed according to an embodiment of the present application; Figure 6 is a working state schematic diagram of the bracket according to an embodiment of the present application; Figure 7 is a structural schematic diagram of an ionization unit according to an embodiment of the present application; Figure 8 is a structural schematic diagram of a fixing seat and a rotating module according to an embodiment of the present application; Figure 9 is a sectional view schematic diagram of the fixing seat and the rotating module according to an embodiment of the present application; Figure 10 is a structural schematic diagram of a moving platform according to an embodiment of the present application; Figure 11 is a structural schematic diagram of a grabbing unit according to an embodiment of the present application; Figure 12 is another state schematic diagram of the grabbing unit according to an embodiment of the present application; Figure 13 is a structural schematic diagram of a moving piece according to an embodiment of the present application; Figure 14 is a structural schematic diagram of a deformation piece according to an embodiment of the present application; Figure 15Figure 6 is a schematic view of a lost material state of the fixed seat and rotating module according to an embodiment of the present application.

[0011] Figures 1-15 The following description and drawings are included to teach any person skilled in the art to whom the present application pertains, how to make and use the same. Some conventional aspects have been simplified or omitted for the sake of clarity. Those skilled in the art will appreciate the changes and alternatives made to the specific embodiments described herein. It will be apparent to those skilled in the art that features described below can be combined in a variety of ways to form numerous variations of the present application. Thus, the present application is not to be limited to the specific embodiments described below, but only by the claims and their equivalents.

[0012] Embodiment 1

[0013] The mass spectrometry-based single-cell analysis system according to Embodiment 1 of the present application, as shown in Figure 1 includes: a transfer station 7 arranged on the base for temporarily storing the electrospray module 11; a three-axis electric claw 4 for grabbing the electrospray module 11 on the support 6 and placing it in the transfer station 7; a grabbing unit 5 for grabbing the electrospray module 11 in the transfer station 7 and moving to the extraction platform 1 for cell extraction, and then placing the electrospray module 11 after cell extraction on the ionization unit.

[0014] As shown in Figure 2 the visualization extraction platform 1 includes a motorized stage 101 for carrying the cell container and providing two-dimensional movement for the cell container, and an imaging unit 104 for obtaining images of the cells in the cell container to select appropriate cells for extraction.

[0015] As shown in Figure 3 the support 6 includes: a second carrying portion 63 having a plurality of parallelly arranged carrying positions 64 for carrying the electrospray module 11; a protection portion 65 arranged on the lower side of the second carrying portion 63, which shields the lower end of the capillary 12 of the electrospray module 11 when the electrospray module 11 is in the carrying position 64 to avoid accidental contact; a connecting portion 66 arranged on the lower side of the protection portion 65 and allowing fixed connection with the base 8.

[0016] As shown in Figure 4As shown, the electrospray module 11 includes a capillary 12, a base 13, and a conductive probe 16, the capillary 12 is inserted into the base 13, the outer diameter of the base 13 is larger than that of the capillary 12.

[0017] In order to quickly and accurately install the electrospray module 11, further, as shown in Figure 5 As shown, the bearing position 64 includes a transverse opening 641 allowing the capillary 12 to pass through, and a longitudinal channel 642 in which the electrospray module 11 is located, and the base 13 is blocked.

[0018] In order to firmly connect for accurate positioning, further, as shown in Figure 6 As shown, the base 8 has a fixed groove allowing the connecting part 66 to be clamped in, and the bottom of the fixed groove has a ball plunger 87, and the bottom side of the mounting part 66 has a groove allowing the ball to be clamped in.

[0019] In order to firmly connect for accurate positioning, further, as shown in Figure 6 As shown, the single-cell analysis system based on mass spectrometry technology further includes: A magnetic plate 67 is arranged on the bracket 1, and a magnet 88 corresponding to the magnetic plate 67 is arranged on the base 8.

[0020] In order to bear and three-dimensionally adjust the position of the electrospray module 11 so that the outlet of the capillary 12 corresponds to the inlet of the mass spectrometer, further, as shown in Figure 7 As shown, the ionization unit 2 includes: The moving platform 50 is used to provide three-dimensional movement, such as vertical direction, left-right direction, and front-back direction; The fixed seat 31 is arranged on the moving platform 50; As shown in Figure 8 The first gear 34 is arranged on the fixed seat 31 and is driven by the first motor 32; The second gear 35 is fixed on the first rotating module 36 and engages with the first gear 34; the first rotating module 36 is arranged on the fixed seat 31; The bearing module 37 is arranged on the first rotating module 36 and is used to bear the electrospray module 11; The second motor 33 and the second rotating module 38, the second rotating module 38 is arranged on the first rotating module 36, and the conductive body 39 is arranged on the second rotating module 38, when the second motor 33 drives the second rotating module 38 to rotate, the conductive body 39 contacts the conductive probe 16.

[0021] In order to automatically place the electrospray module 11, further, the bearing module 37 includes: a groove having an upper and lower portion, the upper portion having a larger width than the lower portion, the lower portion having an opening allowing the capillary tube 12 to pass through, the opening having a width smaller than the outer diameter of the base 13; an extension 373, each side of the upper portion 372 having an upwardly extending portion 373, the distance between the two extensions 373 being not less than the outer diameter of the smaller portion 131 of the base 13 and less than the outer diameter of the larger portion 132 of the base 13.

[0022] To position the electrospray module 11, further as shown in Figure 4 the electrospray module 11 further comprises a carbon steel tube 14 disposed within the base 13; as shown in Figure 9 a magnet 19 for attracting the carbon steel tube 14 is disposed on the fixed seat 31.

[0023] To provide three-dimensional movement with a simple structure, as shown in Figure 10 the movement platform 50 comprises: a base plate 91 disposed on the first guide rail 51 and driven by a motor to translate along the first guide rail 51 in a forward or reverse direction; a second guide rail 52 disposed on the base plate 91 and a third guide rail 53 disposed on the second guide rail 52; a carrier plate 54 disposed on the third guide rail 53 and the fixed seat 31 disposed on the carrier plate 54; a first driving wheel 61 and a second driving wheel 62 driven by a motor and disposed on the base plate 91 on both sides of the second guide rail 52; a plurality of driven wheels, a first driven wheel 71 and a second driven wheel 72 fixed to the base plate 91 on both sides of the second guide rail 52, a third driven wheel 73 and a fourth driven wheel 74 fixed to one side of the third guide rail 53 and on one side of the second guide rail 52, and a fifth driven wheel 75 and a sixth driven wheel 76 fixed to the other side of the third guide rail 53; a synchronous belt 81 passing through the first driving wheel 61, the first driven wheel 71, the fourth driven wheel 74, the sixth driven wheel 76, the second driven wheel 72, the second driving wheel 62, the fifth driven wheel 75, the third driven wheel 73, and the first driving wheel 61 in sequence, the carrier plate 54 connected to the synchronous belt 81 between the fourth driven wheel 74 and the fifth driven wheel 75.

[0024] To recycle used electrospray modules 11, further as shown in Figure 7 a recycling bin 82 having an open upper end and disposed on the base plate 91.

[0025] In order to automate the grabbing of the electrospray module 11, further, as shown in Figures 11-12 the grabbing unit 5 comprises: a fixed support 21 arranged on a three-dimensional mechanical arm; a guide rail 22 and a driving module 85 arranged on the fixed support 21; a moving piece 41 arranged on the guide rail 22 and moving forward or backward along the guide rail 22 under the driving of the driving module 85; As shown in Figure 13 the moving piece 41 comprises an opening 434 and a first bearing part for bearing the electrospray module 11 in communication with the opening 434, the first bearing part comprises an inner diameter decreasing section 431, an inner diameter larger section 432 and an inner diameter smaller section 433 arranged in sequence, the opening 434 allows the capillary 12 to pass through, the inner diameter larger section 432 allows the base 13 to enter, and the inner diameter smaller section 433 allows the capillary 12 to pass through and blocks the base 13; the fixed support 21 has a first gas passage, and the capillary 12 of the electrospray module 11 at the first bearing part is in communication with the first gas passage.

[0026] In order to facilitate the grabbing of the electrospray module 11, further, as shown in Figure 13 the moving piece 41 comprises a first part 42 arranged on the guide rail 22 and a second part 43 connected to the first part 42 and perpendicular to the first part 42, and the second bearing part is arranged on the second part 43.

[0027] In order to realize sealing to improve the ventilation effect, further, the electrospray module 11 further comprises: a deforming piece 15, as shown in Figure 14 the deforming piece 15 is arranged on the upper side of the base 13 and comprises an annular part 152, a body 151 and a cylindrical part 153, the body 151 has a second gas passage 154 inside, the second gas passage 154 has a guide hole 155 allowing the conductive probe 16 to pass through, the annular part 152 is at one end of the body 151, and the cylindrical part 153 is at the other end of the body 151.

[0028] As shown in Figure 4 the cylindrical part 153 is clamped into the base 13, the body 151 and the annular part 152 are exposed outside the base 13, the capillary 12 passes through the base 13, the cylindrical part 153 and the body 151 and is in communication with the second gas passage 154, and the conductive probe 16 passes through the guide hole 155 and enters the capillary 12.

[0029] To improve the positioning accuracy and sealing effect, further, the fixed support 21 has an annular groove around the outlet of the first gas passage 152, which allows the annular part to enter, and the central axis of the annular groove is collinear with the central axis of the first bearing part.

[0030] To improve the sealing effect between the deformed part 15 and the base 13, further, the outer side of the cylindrical part 153 has a plurality of annular protrusions 156 which are clamped into the base 13.

[0031] The single-cell mass spectrometry method of the embodiment of the application, that is, the working method of the single-cell analysis system based on mass spectrometry technology of the embodiment, comprises the following steps: (A1) The three-axis electric claw 4 grabs the electrospray module 11 on the support 6 and places it on the transfer site 7; (A2) The grabbing unit 5 grabs the electrospray module 11 at the transfer site 7 and moves to the extraction platform 1; (A3) The capillary tube 12 of the electrospray module 11 completes the visualization of the selected cells on the extraction platform 1, and the capillary tube 12 absorbs the extraction material; (A4) The grabbing unit 5 continues to move the electrospray module 11 and places it on the ionization unit 2; (A5) The ionization unit 2 adjusts the position of the electrospray module 11 through translation and rotation, so that the outlet of the capillary tube 12 corresponds to the sample inlet cone of the mass spectrometer; (A6) The electrically conductive probe 16 in the capillary tube 12 discharges, the extraction material is ionized, and the ions enter the sample inlet cone.

[0032] In the above step (A1), the feeding of the electrospray module 11 is completed first, specifically: As shown in Figure 5 , a plurality of electrospray modules 11 are respectively arranged in an inclined manner in each bearing position 64 of the support 1, and then the electrospray modules 11 are adjusted to a vertical state; A downward pressure is applied to the electrospray module 11, so that the electrospray module 11 is vertically arranged in the bearing position 64, and the lower end of the capillary tube 12 is protected by the protection part 65, as shown in Figure 3 ; The support 1 is fixed on the base of the single-cell analysis system based on mass spectrometry technology, as shown in Figure 3 , Figure 6 .

[0033] To quickly and accurately set the electrospray module 11, further, as shown in Figure 5As shown, in step (A1), the capillary tube 12 passes through the transverse opening 641 of the bearing site 64, and the base 13 is inserted into the longitudinal channel 642 of the bearing site 64; The rotary electrospray module 11 is shown, with the capillary tube 12 passing through the transverse opening 641 and vertically located in the longitudinal channel 642; In the downward movement, the base 13 is blocked, and the lower end of the capillary tube 12 is shielded by the front and rear protection parts 65, i.e., protected from accidental contact.

[0034] In step (A5), the specific adjustment method is as follows: The electrospray module 11 is located in the bearing module 37, which is arranged on the first rotary module 36, and the first rotary module 36 is arranged on the fixed seat 31; The second rotary module 38 is driven to rotate in the forward direction, so that the conductive body 39 on the second rotary module 38 contacts the conductive probe 16 in the electrospray module 11; The moving platform 50 drives the fixed seat 31, so that the outlet of the capillary tube 12 of the electrospray module 11 is located at the sampling orifice, the conductive probe 16 discharges to form an electrospray, the substance in the capillary tube 12 is ionized, and the ions enter the sampling orifice; After ionization is completed, the moving platform 50 drives the fixed seat 31 to reach the upper side of the collection tank 82; The second rotary module 38 is driven to rotate in the reverse direction, and the conductive body 39 is separated from the conductive probe 16; The first gear 34 on the fixed seat 31 rotates to drive the second gear 35 on the first rotary module 36, so that the first rotary module 36 rotates, and the bearing module 37 tilts, i.e., the smaller width part 371 of the groove is on the upper side, and the larger width part 372 is on the lower side, as shown in Figure 11 As shown, the electrospray module 11 automatically slides into the collection tank 82 to achieve automatic recovery.

[0035] In order to automatically place the electrospray module 11, further, the method for placing the electrospray module 11 into the ionization unit 2 is as follows: The electrospray module 11 is fixed on the moving part of the grabbing unit, the mechanical arm moves the grabbing unit above the obliquely arranged bearing module 37, and then moves downward, so that the smaller outer diameter part 131 of the base 13 of the electrospray module 11 enters between the two extension parts 373, and the larger outer diameter part 132 of the base 13 is located on the upper side of the extension part 373; The driving module 85 of the grabbing unit drives the moving part 41 to move downward, and the larger outer diameter part 132 is blocked by the extension part 373 until the electrospray module 11 is separated from the moving part 41; The electrospray module 11 falls from between the two extension portions 373, enters the groove of the carrier module 37, and moves downward along the groove, with the capillary 12 passing through the opening on the lower side of the groove.

[0036] For accurate positioning, a ferrous round tube 14 is arranged in the base 13, and a magnet 19 is arranged on the fixing seat 31. The electrospray module 11 in the carrier module 37 is positioned by the adsorption force of the magnet 19 and the carbon steel round tube 14.

[0037] The manner of grabbing the electrospray module 11 by the grabbing unit 5 is as follows: Under the driving of the driving module 85, the moving piece 41 moves forward along the guide rail 22 arranged on the fixing support 21; As shown in Figure 11 The moving piece 41 is moved, the capillary 12 passes through the opening 434 on the moving piece 41 and enters the smaller inner diameter section 433, and the base 13 enters the larger inner diameter section 432 and is blocked by the smaller inner diameter section 433; As shown in Figure 12 Under the driving of the driving module 85, the moving piece 41 moves reversely along the guide rail 22, one end of the electrospray module 11 contacts and seals the fixing support 21, and the first gas passage in the fixing support 21 communicates with the capillary 12.

[0038] Embodiment 2: Application example of the single-cell analysis system and method based on mass spectrometry technology according to Embodiment 1 of the present application.

[0039] In this application example, as shown in Figure 1 A plurality of racks 6 are fixed side by side on the base, and 23 brand-new (as consumables) electrospray modules 11 are arranged on each rack 6. The racks 6, the transfer site 7, the visual extraction platform 1, and the ionization unit 2 are arranged in sequence in the left-right direction.

[0040] The three-axis electric claw 4 is used to grab the electrospray module 11 on the rack 6 and transfer it to the transfer site. The three-axis electric claw 4 is prior art in the field.

[0041] As shown in Figure 2 In the visual extraction platform, the motorized stage 101 moves in two dimensions in the horizontal plane, so as to adjust the relative position of the carried cell container and the objective 102. The lower side of the motorized stage 101 is transparent, the objective 102 is arranged on the lower side of the motorized stage 101, the objective 1, the lens 105, and the camera 104 are arranged in sequence on the optical path, and the optical path is arranged in the lens barrel 103. By imaging and two-dimensional movement, the cells are selected, and then the electrospray module 11 grabbed by the grabbing unit 5 completes the extraction.

[0042] AsFigure 3 As shown, 23 carrying positions 64 are arranged in parallel on the second carrying part 63, i.e. 23 electrospray modules 11 are arranged at most. The protection part 65 is provided with two strip-shaped blocking parts arranged at the front and back of the bracket 1 respectively. The two ends of the second carrying part 63 and the protection part 65 are connected with the vertical part 68, and the bottom end of the vertical part 68 is fixed on the connecting part 66. The handle 69 is arranged on one side of the vertical part 68. The magnetic plate 67 is L-shaped and fixed on the upper side of the connecting part 66 and adjacent to the vertical part 68.

[0043] As shown in the drawing, Figure 4 In the electrospray module 11, the base 13 includes a larger-diameter part 132 and a smaller-diameter part 131. The cylindrical part 153 is inserted into the base 13 to press the carbon steel pipe 14 in the base 13, and the body 151 and the annular part 152 are exposed outside the base 13. The capillary tube 12 passes through the base 13, the cylindrical part 153 and the body 151 and communicates with the second gas passage 154, and the center axis of the guide hole 155 is collinear with the center axis of the capillary tube 12. The conductive probe 15 passes through the guide hole 155 and enters the capillary tube 12, and the spherical conductive body 17 is fixed on one end of the conductive probe 16 and located in the annular part 152.

[0044] As shown in the drawing, Figure 5 The carrying position 64 includes a transverse opening 641 and a longitudinal passage 642. The transverse opening 641 allows the capillary tube 12 to pass through but prevents the base 13 from passing through. The longitudinal passage 642 is provided with a blocking part to block the base 13 from passing through, so that the electrospray module 11 is vertically arranged in the longitudinal passage 642.

[0045] As shown in the drawing, Figure 6 When the bracket 1 is fixed in the fixing groove of the base 8, the plurality of ball plunger 87 on the base is inserted into the groove on the lower side of the connecting part, and the magnetic plate 67 and the magnet 88 on the base 8 are attracted, which ensures the accurate positioning of the electrospray module 11 on the bracket 1.

[0046] As shown in the drawing, Figure 7 In the ionization unit 2, the first guide rail 51 is vertically arranged, the two second guide rails 52 are horizontally arranged in front and back, and the two third guide rails 53 are horizontally arranged left and right. The fixing seat 31 is arranged on the carrying plate 54, and the carrying plate 54 is arranged on the third guide rail 53 and driven to translate along the third guide rail 53.

[0047] As shown in the drawing, Figures 8-9As shown, in the ionization unit, a magnet is provided on the fixed base 31 to attract the carbon steel round tube 14 of the electrospray module 11 located in the carrier module 37. A first motor 32 is provided on the fixed base 31 to drive the first gear 34 to rotate. A first rotating module 36 is provided on the fixed base 31, and a second gear 35 is fixed on the first rotating module 36. When the first gear 34 with a smaller outer diameter drives the second gear with a larger outer diameter, the first rotating module 36 rotates in place around the central axis of the second gear 35.

[0048] The support module 37 is fixed to the first rotating module 36. The groove includes a wider portion 372 that accommodates the larger outer diameter portion 132 of the base 13 and a narrower portion 371 that accommodates the smaller outer diameter portion 131 of the base 13. The end of the narrower portion 371 has an opening that allows the capillary 12 to pass through, and this opening blocks the base 13. The two sides of the wider portion 372 each have an extension portion 373. The distance between the two extension portions 373 is not less than the outer diameter of the narrower outer diameter portion 131 of the base 13, but less than the outer diameter of the wider portion 132, thus preventing the electrospray module 11 from completely passing through.

[0049] The second motor 33 is mounted on the first rotating module 36, and the second rotating module 38 is mounted on the first rotating module 36. When the second motor 33 drives the second rotating module 38 to rotate, the conductor 39 contacts and disconnects from the conductive probe 16 of the electrospray module 11.

[0050] like Figure 10 As shown, in the mobile platform 50, the first driving wheel 61, the second driving wheel 62, the first driven wheel 71, and the second driven wheel 72 are respectively fixed on the base plate 91. The first driving wheel 61 and the first driven wheel 62 are located on the left side of the two second guide rails 52, and the second driving wheel 62 and the second driven wheel 72 are located on the right side of the second guide rails 52. The third driven wheel 73 and the fourth driven wheel 74 are respectively fixedly connected to the front and rear sides of the third guide rail 53 and are located on the left side of the third guide rail 53. The fifth driven wheel 75 and the sixth driven wheel 76 are respectively fixedly connected to the front and rear sides of the third guide rail 53 and are located on the right side of the third guide rail 53. The synchronous belt 81 passes sequentially around the first driving pulley 61, the first driven pulley 71, the fourth driven pulley 74, the sixth driven pulley 76, the second driven pulley 72, the second driving pulley 62, the fifth driven pulley 75, the third driven pulley 73 and the first driving pulley 61, and the bearing plate 54 connects the synchronous belt 81 located between the fourth driven pulley 74 and the fifth driven pulley 75.

[0051] When the first drive wheel 61 rotates counterclockwise and the second drive wheel 62 rotates clockwise, the timing belt 81 drags the third guide rail 53 to move forward along the second guide rail 52, which in turn drives the bearing plate 54 and the fixed seat 31 to move forward. When the first driving wheel 61 rotates clockwise and the second driving wheel 62 rotates counterclockwise, the synchronous belt 81 drags the third guide rail 53 to move backward along the second guide rail 52, that is, drags the bearing plate 54 and the fixed seat 31 to move backward; When the first driving wheel 61 and the second driving wheel 62 both rotate counterclockwise, the synchronous belt 81 drags the bearing plate 54 to move left along the third guide rail 53; When the first driving wheel 61 and the second driving wheel 62 both rotate clockwise, the synchronous belt 81 drags the bearing plate 54 to move right along the third guide rail 53.

[0052] As shown in Figures 11-12 The fixed support 21 is provided with a linear guide rail 22. The driving module 85 includes a push rod electric motor 86 and a movable push rod 87, and the movable push rod 87 is connected to the first part 42 of the moving part 41.

[0053] As shown in Figure 13 The first part 42 and the second part 43 are vertically arranged, and the first part 42 is arranged on the guide rail 22. The second part 43 is provided with an opening 434 and a second bearing part, and the second bearing part includes sequentially arranged an inner diameter decreasing section 431 (convenient for the base 13 to enter the inner diameter larger section 432), an inner diameter larger section 432 and an inner diameter smaller section 433. The inner diameter larger section 432 is matched with the base 13 of the electrospray module 11 (the difference between the inner diameter of the inner diameter larger section 432 and the outer diameter of the base 13 is small), and the inner diameter of the inner diameter smaller section 432 is larger than the outer diameter of the capillary 12 but smaller than the outer diameter of the base 13, so that the inner diameter smaller section 433 blocks the base 13, that is, limits the axial movement of the ion source 11, and the inner diameter larger section 432 limits the radial movement of the electrospray module 11. The width of the opening 434 is larger than the outer diameter of the capillary 12 but smaller than the outer diameter of the base 13, that is, allows the capillary 12 to pass through but blocks the base 13.

[0054] As shown in Figure 14 The deforming part 15 is made of silica gel, and the body 151 is internally provided with a second gas passage 154, the second gas passage 154 is internally provided with a guide hole 155 allowing the conductive probe 16 to pass through, the annular part 152 is located at one end of the body 151, the cylindrical part 153 is located at the other end of the body 151, a plurality of annular protrusions 156 are located outside the cylindrical part 153, and the lower end opening 157 of the cylindrical part 153 is trumpet-shaped, facilitating the insertion of the capillary 12. The fixed support 21 is internally provided with a first gas passage, and an annular groove surrounding the opening of the first gas passage is arranged on the sidewall of the fixed support 21, and the annular groove allows the annular part 152 to be clamped in, achieving sealing.

[0055] The mass spectrometric analysis method of the embodiment, that is, the working method of the single-cell analysis system based on mass spectrometry of the embodiment, includes the following steps: (A1) Feeding of electro-spray module 11, specifically: like Figure 5 As shown, multiple electrospray modules 11 are obliquely arranged at various bearing positions 64 of the bracket 1. Specifically, the capillary tube 12 passes through the transverse opening 641 of the bearing position 64, and the base 13 is inserted into the longitudinal channel 642 of the bearing position 64. The rotating electro-spray module 11 has its capillary 12 extending beyond the transverse opening 641 and vertically positioned within the longitudinal channel 642.

[0056] A downward pressure is applied to the electro-spray module 11, causing it to be vertically positioned in the bearing position 64. The lower end of the capillary 12 is shielded by the protective part 65 to prevent accidental contact. Figure 3 As shown; The scaffold 1 is fixed to the base of the single-cell analysis system based on mass spectrometry technology, specifically as follows: Pushing handle 69 causes connecting part 66 to enter the fixing groove on base 8. Ball-head plunger 87 on base engages with the groove on the lower side of connecting part 66. Magnetic plate 67 attracts magnet 88 on base 8, thus accurately and securely fixing bracket 1 to base 8. Figure 6 As shown.

[0057] During downward movement, the base 13 is blocked; the lower end of the capillary 12 is shielded by the front and rear protective parts 65 respectively, thus protecting it from accidental contact.

[0058] The three-axis electric gripper 4 grasps the electro-spray module 11 on the bracket 6 and places it in the transfer position 7; (A2) The gripping unit 5 grips the electrospray module 11 at the intermediate transfer position 7 and moves it to the extraction platform 1, specifically in the following manner: Driven by the drive module 85, the moving part 41 moves forward along the guide rail 22; like Figure 11 As shown, the three-dimensional robotic arm moves and grasps the unit, so that the capillary 12 of the electro-spray module 11 at the intermediate position 7 passes through the opening 434 on the moving part 41 and enters the section with a smaller inner diameter 433 and the section with a larger inner diameter 432. like Figure 12 As shown, under the drive of the drive module 85, the moving part 41 moves in the opposite direction along the guide rail 22, so that the base 13 enters the section with a larger inner diameter 432 and is blocked by the section with a smaller inner diameter 433. Subsequently, the annular portion 152 at one end of the electrospray module 11 enters the annular groove on the side wall of the fixed support and is deformed by compression, thereby achieving a seal between the fixed support 21 and the electrospray module 11. The first gas channel in the fixed support 21 is connected to the capillary 12 through the second gas channel 154.

[0059] (A3) Visualizing the selected cells on the extraction platform 1 by using the capillary 12 of the electrospray module 11, which sucks the extraction material; the specific extraction method is the same as the prior art.

[0060] (A4) The gripping unit 5 continues to move the electrospray module 11 and places it on the ionization unit 2 in the following way: The three-dimensional mechanical arm moves the gripping unit 5 so that the gripping unit 5 is obliquely above the carrying module 37, at which time the carrying module 37 is obliquely arranged; The gripping unit moves downward so that the smaller outer diameter part 131 of the base 13 of the electrospray module 11 enters between the two extension parts 373, and the larger outer diameter part 132 of the base 13 is obliquely above the extension parts 373; The driving module 85 drives the moving part 41 to move obliquely downward, and the larger outer diameter part 132 is blocked by the extension parts 373 until the electrospray module 11 is separated from the moving part 41; The electrospray module 11 falls by itself from between the two extension parts 373 into the groove of the carrying module 37 and moves downward along the inclined groove, the capillary 12 passes through the opening on the lower side of the groove, the smaller outer diameter part 131 is in the smaller width part 371, and the larger outer diameter part 132 is in the larger width part 372, and at the same time, under the adsorption of the magnet 19, the electrospray module 11 is fixed in the carrying module 37.

[0061] (A5) The ionization unit 2 adjusts the position of the electrospray module 11 by translation and rotation so that the outlet of the capillary 12 corresponds to the sampling orifice of the mass spectrometer in the following way: The second motor 33 drives the second rotating module 38 to rotate in the positive direction around its rotation shaft 302 so that the conductive body 39 on the second rotating module 38 contacts the conductive probe 16 in the electrospray module 11; (S3) The moving platform 50 drives the fixing seat 31 so that the outlet of the capillary 12 of the electrospray module 11 is accurately at the sampling orifice under the monitoring of the camera on the fixing seat 31.

[0062] (A6) The conductive probe 16 in the capillary 12 discharges, the extraction material is ionized, and the ions enter the sampling orifice.

[0063] After the ionization is completed, the moving platform 50 drives the fixing seat 31 to the upper side of the collection box 82; The second motor 33 drives the second rotating module 38 to rotate in the reverse direction, and the conductive body 39 is separated from the conductive probe 16 to avoid blocking the ion source 11 when it is recycled; The first gear 34 on the fixed seat 31 rotates to drive the second gear 35 on the first rotating module 36, so that the first rotating module 36 rotates in situ around its rotating shaft 301 (the central axis of the rotating shaft 301 and the central axis of the second gear 35 are collinear), and the bearing module 37 is tilted, that is, the part 371 with a smaller width of the groove is on the top, and the part 372 with a larger width of the groove is on the bottom, as shown in Figure 15 The electrospray module 11 automatically slides into the groove and falls into the collection box 82 to realize automatic recovery.

[0064] In the above process, the mobile platform 50 works as follows: When the first driving wheel 61 rotates clockwise and the second driving wheel 62 rotates counterclockwise, the synchronous belt 81 drags the third guide rail 53 to move backward along the second guide rail 52, that is, the bearing plate 54 and the fixed seat 31 are moved backward; When the first driving wheel 61 and the second driving wheel 62 both rotate counterclockwise, the synchronous belt 81 drags the bearing plate 54 to move left along the third guide rail 53; When the first driving wheel 61 and the second driving wheel 62 both rotate clockwise, the synchronous belt 81 drags the bearing plate 54 to move right along the third guide rail 53.

[0065] According to the technical scheme of the embodiment, the grabbing, placing, three-dimensional movement, electrical conduction, ionization and recovery of the electrospray module 11 are all realized automatically, without manual operation, and the efficiency of single-cell analysis is significantly improved.

Claims

1. A single cell analysis system based on mass spectrometry technology, comprising a support, a plurality of electrospray modules arranged on the support, an extraction platform and a mass spectrometer arranged on a base; the electrospray module comprises a base, a conductive probe and a capillary tube, the capillary tube is inserted into the base, the outer diameter of the base is larger than the outer diameter of the capillary tube, and the conductive probe is inserted into the capillary tube; characterized in that, The single-cell analysis system further comprises: a transfer position for temporarily storing the electrospray module; a three-axis electric claw for grabbing the electrospray module on the support and transferring to the transfer position; a grabbing unit for grabbing the electrospray module in the transfer position and moving to an extraction platform for cell extraction, and then placing the electrospray module after cell extraction on an ionization unit; the ionization unit for carrying the electrospray module and adjusting its position, and the outlet of the capillary corresponds to the sampling orifice of the mass spectrometer.

2. The mass spectrometry-based single-cell analysis system of claim 1, wherein, The extraction platform comprises: an electric carrying stage for carrying the cell container and providing two-dimensional movement for the cell container; an imaging unit for obtaining images of the cells in the cell container.

3. The mass spectrometry-based single-cell analysis system of claim 1, wherein, The grabbing unit comprises: a fixed support and a driving module arranged on the fixed support; a moving part that moves forward or reversely under the driving of the driving module; the moving part comprises an opening and a first carrying part for carrying the electrospray module in communication with the opening, the first carrying part comprises a section with gradually decreasing inner diameter, a section with relatively large inner diameter and a section with relatively small inner diameter arranged in sequence, the opening allows the capillary to pass through, the section with relatively large inner diameter allows the base to enter, and the section with relatively small inner diameter allows the capillary to pass through and blocks the base; the fixed support has a first gas passage, and the capillary of the consumable in the first carrying part communicates with the first gas passage.

4. The mass spectrometry-based single cell analysis system of claim 1, wherein, The electrospray module further comprises: a deformation part comprising a ring-shaped part, a body and a cylindrical part, the body has a second gas passage inside, the second gas passage has a guide hole allowing the conductive probe to pass through, the ring-shaped part is at one end of the body, the cylindrical part is at the other end of the body, the cylindrical part is clamped into the base, the capillary passes through the base, the cylindrical part and the body and communicates with the second gas passage, and the conductive probe passes through the guide hole and enters the capillary.

5. The mass spectrometry-based single-cell analysis system of claim 1, wherein, The ionization unit comprises: a fixed seat arranged on a moving platform; a first motor and a first gear arranged on the fixed seat and driven by the first motor; a first rotating module and a second gear fixed on the first rotating module and engaged with the first gear; the first rotating module is arranged on the fixed seat; a carrying module fixed on the first rotating module for carrying the electrospray module; a second motor and a second rotating module arranged on the first rotating module, and a conductive body arranged on the second rotating module; when the second motor drives the second rotating module to rotate, the conductive body contacts the conductive probe.

6. The mass spectrometry-based single-cell analysis system of claim 5, wherein, The carrying module comprises: a groove having an opening on the lower side allowing the capillary to pass through, the width of the opening is smaller than the outer diameter of the base, and the groove has a larger width part and a smaller width part arranged in sequence from top to bottom; The two sides of the larger-width part of the base have upward extensions, the distance between the two extensions is not less than the outer diameter of the smaller-width part and is less than the outer diameter of the larger-width part.

7. The mass spectrometry-based single-cell analysis system of claim 1, wherein, The support includes: The second bearing part has a plurality of bearing positions arranged side by side, and the bearing positions are used to bear the electrospray module; The protection part is located at the lower side of the second bearing part, and protects the lower end of the capillary when the electrospray module is in the bearing position; the two ends of the second bearing part and the protection part are connected to the support; The connecting part is located at the lower side of the protection part and allows fixed connection with the base.

8. The mass spectrometry-based single-cell analysis system of claim 7, wherein, The bearing position includes a transverse opening and a longitudinal channel, the opening allows the capillary to pass through, and the electrospray module is in the longitudinal channel, and the base is blocked.

9. The single-cell analysis method of the single-cell analysis system according to any one of claims 1-8, comprising the following steps: (A1) The three-axis electric claw grabs the electrospray module on the support and places it in the transfer position; (A2) The grabbing unit grabs the electrospray module at the transfer position and moves to the extraction platform; (A3) The capillary of the electrospray module completes the visualization of the selected cells on the extraction platform, and the capillary absorbs the extraction material; (A4) The grabbing unit continues to move the electrospray module and places it on the ionization unit; (A5) The ionization unit adjusts the position of the electrospray module by translation and rotation, so that the outlet of the capillary corresponds to the sampling cone of the mass spectrometer; (A6) The conductive probe in the capillary discharges, the extraction material is ionized, and the ions enter the sampling cone.

10. The single-cell analysis method according to claim 9, wherein, In step (A5), the specific adjustment method is: The electrospray module is in the bearing module, the bearing module is arranged on the first rotating module, and the first rotating module is arranged on the fixed base; The second rotating module is driven to rotate forward, so that the conductive body on the second rotating module contacts the conductive probe in the electrospray module; The moving platform drives the fixed base, so that the outlet of the capillary of the electrospray module is at the sampling cone, the conductive probe discharges, forms an electrospray, the material in the capillary is ionized, and the ions enter the sampling cone; After ionization is completed, the moving platform drives the fixed base to the upper side of the collection box; The second rotating module is driven to rotate reversely, and the conductive body is separated from the conductive probe; The first gear on the fixed base rotates to drive the second gear on the first rotating module, so that the first rotating module rotates, the bearing module tilts, and the electrospray module automatically slides into the collection box.

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