CN115436451A - Single cell mass spectrometry systems and methods - Google Patents
Single cell mass spectrometry systems and methods Download PDFInfo
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- CN115436451A CN115436451A CN202210370626.6A CN202210370626A CN115436451A CN 115436451 A CN115436451 A CN 115436451A CN 202210370626 A CN202210370626 A CN 202210370626A CN 115436451 A CN115436451 A CN 115436451A
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- 238000004949 mass spectrometry Methods 0.000 title claims abstract description 40
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- 238000004458 analytical method Methods 0.000 claims abstract description 8
- 239000004020 conductor Substances 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 9
- 238000001819 mass spectrum Methods 0.000 claims description 5
- 238000007789 sealing Methods 0.000 claims description 4
- 238000004891 communication Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 description 5
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Abstract
The invention provides a single-cell mass spectrometry system and a single-cell mass spectrometry method, wherein the single-cell mass spectrometry system comprises a capillary needle, a single-cell bearing device and a mass spectrometer; the capillary needle comprises a first part, a second part and a third part which are sequentially distributed, wherein the outer diameter of the second part is gradually increased; the capillary needle is arranged on the moving piece, and the moving piece is used for moving the capillary needle to the single cell bearing device, the mass spectrometer and the capillary needle bearing device respectively; the capillary needle bearing device is provided with a plurality of capillary needle bearing positions; the clamping device comprises a base, a first clamping jaw, a first guide rail, a driving unit and a second clamping jaw, wherein the base is fixed on a bearing piece; the first clamping jaw and the second clamping jaw are respectively provided with oppositely arranged grooves, and the distance between the bottom walls of the two grooves is increased from the lower walls of the grooves to the upper part; the second portion and the third portion are clamped within the two recesses of the first jaw and the second jaw. The invention has the advantages of accurate analysis and the like.
Description
Technical Field
The invention relates to single cell analysis, in particular to a single cell mass spectrometry system and a single cell mass spectrometry method.
Background
The cell is an important research object for human to research life secret and explain disease mechanism. Traditional cell biology studies often target population cells, and have explored the pathogenesis and therapeutic approaches of many diseases for humans. But there are significant drawbacks based on the average study of the population cells, ignoring cellular "heterogeneity". The single cell analysis can make up for the important information which is covered and omitted in the past due to the sampling of the group cells, so that the research of human beings on diseases is more accurate and comprehensive. The single cell mass spectrometry technology can realize simultaneous detection of multi-component samples, obtain structural information of unknown components in a large number of samples, has higher sensitivity, and provides effective technical support for researches of single cell metabonomics, proteomics and the like.
The defects of the current single-cell mass spectrometry technology mainly lie in that:
1. the capillary needle runs through the whole process for use, so that the potential risk of cross contamination is high, and the scheme based on the capillary needle has the problem of cross contamination among single cells;
the capillary needle cleaning technology is introduced into part of schemes, but the system work flow becomes more complicated, and the reliability is reduced.
2. The bilateral capillary needles are operated cooperatively, even the intervention of a rotating motor is possibly required, the control is complex, and the error is easy to occur.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a single-cell mass spectrometry system.
The purpose of the invention is realized by the following technical scheme:
the single-cell mass spectrometry system comprises a capillary needle, a single-cell bearing device and a mass spectrum analyzer; the capillary needle comprises a first part with smaller outer diameter, a second part with gradually larger outer diameter and a third part with larger outer diameter which are distributed in sequence; the single-cell mass spectrometry system further comprises:
the bearing part is arranged on the moving part, the capillary needle is arranged at one end of the bearing part, which is far away from the moving part, and the moving part is used for moving the capillary needle to the single cell bearing device, the mass spectrometer and the capillary needle bearing device respectively;
the capillary needle carrying device is provided with a plurality of carrying positions suitable for carrying the capillary needles;
the clamping device comprises a base, a first clamping jaw, a second clamping jaw, a first guide rail and a driving unit, wherein the base is fixed on the bearing piece, the first clamping jaw is arranged on the base, the first guide rail is arranged on the base, and the driving unit is used for driving the second clamping jaw to move on the first guide rail in the positive direction and the direction; the first clamping jaw and the second clamping jaw are respectively provided with oppositely arranged grooves, and the distance between the side walls of the two grooves is increased from the lower walls of the grooves to the upper part; the second part and the third part are clamped in the two grooves of the first clamping jaw and the second clamping jaw.
Another object of the present invention is to provide a single-cell mass spectrometry method according to the above single-cell mass spectrometry system, and the object of the present invention is achieved by the following technical solutions:
according to the single-cell mass spectrometry method of the single-cell mass spectrometry system, the single-cell mass spectrometry method comprises the following steps:
(A1) The moving piece moves the bearing piece to the capillary needle bearing device, and the clamping device moves to the selected capillary needle;
(A2) The driving unit drives the second clamping jaw to move forwards on the first guide rail, and the third part and the second part of the capillary needle enter the two grooves and are clamped by the first clamping jaw and the second clamping jaw.
Compared with the prior art, the invention has the following beneficial effects:
1. cross contamination is avoided;
the capillary needle bearing device is provided with a plurality of capillary needles which are distributed in a matrix form, in each single cell detection, a new capillary needle is firstly clamped and then used for single cell extraction and detection, and the capillary needles are used as consumables, so that the cross contamination among the capillary needles is avoided;
2. the structure is simple;
at the same time, only one capillary needle moves along with the moving part, and the two-dimensional moving platform is arranged on the moving part, namely only one three-dimensional moving device (the combination of the moving part and the two-dimensional moving platform) is needed to realize the three-dimensional movement of the capillary needle, so that the single cell extraction and detection are realized, other control parts (such as a rotating motor and the like) are not needed, the complexity of the system is reduced, and the reliability is improved;
3. the analysis efficiency is high;
the loading, the single cell extraction and the detection of the capillary needle are automatically realized, so that the single cell analysis efficiency is obviously improved;
the design of the second part and the third part of the capillary needle and the design of the first groove and the second groove (the fourth part and the fifth part) matched with the second part and the third part of the capillary needle enable the capillary needle to be clamped by the first clamping jaw and the second clamping jaw quickly and accurately, and the overall working efficiency is improved.
Drawings
The disclosure of the present invention will become more readily understood with reference to the accompanying drawings. As is readily understood by those skilled in the art: these drawings are only for illustrating the technical solution of the present invention and are not intended to limit the scope of the present invention. In the figure:
FIG. 1 is a schematic diagram of a single-cell mass spectrometry system according to an embodiment of the present invention.
Detailed Description
Fig. 1 and the following description depict alternative embodiments of the invention to teach those skilled in the art how to make and reproduce the invention. Some conventional aspects have been simplified or omitted in order to explain the technical scheme of the present invention. Those skilled in the art will appreciate that variations or substitutions from these embodiments will be within the scope of the invention. Those skilled in the art will appreciate that the features described below can be combined in various ways to form multiple variations of the invention. Thus, the present invention is not limited to the following alternative embodiments, but is only limited by the claims and their equivalents.
Example 1:
fig. 1 is a schematic diagram showing a structure of a single-cell mass spectrometry system according to example 1 of the present invention, and as shown in fig. 1, the single-cell mass spectrometry system includes:
the capillary needle 14, the single cell carrying device 31 and the mass spectrum analyzer 51; the capillary needle 14 comprises a first part with a smaller outer diameter, a second part with a gradually larger outer diameter and a third part with a larger outer diameter which are distributed in sequence;
a moving member 11 and a carrier 12, wherein the carrier 12 is disposed on the moving member 11, the capillary needle 14 is disposed at an end of the carrier 12 far from the moving member 11, and the moving member 11 is configured to move the capillary needle 14 to the single cell holder 41, the mass analyzer 51 and the capillary needle holder 31, respectively, such as moving the moving member 11 by a linear track or a circular track;
a capillary needle carrier 31, wherein the capillary needle carrier 31 has a plurality of carrying positions suitable for carrying the capillary needles 14, for example, a plurality of capillary needles are distributed on the capillary needle carrier 31 in a matrix manner and arranged in parallel;
the clamping device 13 comprises a base, a first clamping jaw, a second clamping jaw, a first guide rail and a driving unit, wherein the base is fixed on the bearing piece, the first clamping jaw is arranged on the base, the first guide rail is arranged on the base, and the driving unit is used for driving the second clamping jaw to move forwards and backwards on the first guide rail; the first clamping jaw and the second clamping jaw are respectively provided with oppositely arranged grooves, and the distance between the side walls of the two grooves is increased from the lower wall of each groove to the upper part of each groove; the second part and the third part are clamped in the two grooves of the first clamping jaw and the second clamping jaw.
To achieve three-dimensional movement of the capillary needle, further, the single-cell mass spectrometry system further comprises:
a second guide rail 21, wherein the moving member 11 is arranged on the second guide rail 21;
the two-dimensional moving platform is arranged on the moving part, and the bearing part is arranged on the two-dimensional moving platform.
In order to realize single cell extraction and ionization, the base is internally provided with a gas channel, the capillary needle is provided with a through hole penetrating through the first part, the second part and the third part, and the capillary is arranged in the through hole; when the capillary needle is positioned in the two grooves, the through hole is communicated with the gas channel, namely, the communication between the gas channel and the capillary tube is realized.
In order to realize the ionization of the substance to be detected, further, the single-cell mass spectrometry system further comprises:
the electric conductor is arranged in the gas channel;
an electrode disposed within the capillary needle; when the capillary needle is in the two grooves, the electric conductor is connected with the electrode.
In order to improve the reliability of the electrical connection, further, the electrode includes a ball portion and a needle portion extending in the capillary; when the capillary needle is positioned in the two grooves, the electric conductor is connected with the spherical part; the first clamping jaw is fixed on the base.
In order to realize air path sealing, the side part of the base, which is adjacent to the capillary needle, is provided with an annular groove which surrounds the opening of the air passage, and the sealing element is arranged in the annular groove; when the capillary needle is positioned in the two grooves, the sealing element is pressed by the base and the capillary needle.
In order to stably and reliably drive the second clamping jaw at a low speed, further, the driving unit comprises:
the motor is fixed on the base and drives the gear to rotate;
the linear connecting piece is arranged on the first guide rail and connected with the second clamping jaw, and the gear is meshed with the teeth on the connecting piece.
The single-cell mass spectrometry method of the embodiment of the invention, that is, the working method of the single-cell mass spectrometry system of the embodiment of the invention, comprises the following steps:
(A1) The moving member 11 moves the carrier 12 to the capillary needle carrier 31, and the holding device 13 moves to the selected capillary needle 14;
(A2) The driving unit drives the second clamping jaw to move forwards on the first guide rail, and the third part and the second part of the capillary needle 14 enter the two grooves and are clamped by the first clamping jaw and the second clamping jaw;
(A3) The moving member 11 moves the capillary needle 14 to the single cell carrying device 41 to complete the extraction of the selected single cell;
(A4) The moving member 11 moves the capillary needle 14 to the mass analyzer 51 to complete the analysis of the single cell;
(A5) The driving unit drives the second clamping jaw to move reversely on the first guide rail, the first clamping jaw is separated from the second clamping jaw, and the capillary needle falls off and is collected.
In order to realize the gas-electric simultaneous conduction, further, in the step (A2), when the capillary needle is clamped by the first clamping jaw and the second clamping jaw, the electric conductor in the gas channel on the base is connected with the electrode in the capillary needle, and meanwhile, the inside of the capillary needle 14 is communicated with the gas channel.
Example 2:
the single-cell mass spectrometry system and the working method thereof according to the embodiment 1 of the invention are applied.
In the present application example, as shown in fig. 1, the second guide rail 21 is a linear guide rail, the moving member 11 is disposed on the second guide rail 21, and the two-dimensional moving platform is disposed on the moving member 11 and combined to form a three-dimensional moving platform; one end of the bearing part 12 is fixed on the two-dimensional moving platform, and the other end is fixed with the clamping device 13; the single cell bearing device 41 is arranged on the electric objective table, and the microscopic imaging module is arranged at the lower side of the single cell bearing device 41;
the capillary needles 14 are arranged on the capillary needle carrying device 31 in a matrix manner, the capillary needles 14 are mutually parallel, and the top end positions of the capillary needles 14 are coplanar; from bottom to top, the capillary needle 14 comprises a first part with a smaller outer diameter, a second part with a gradually larger outer diameter (from the outer diameter of the first part to the outer diameter of the third part), and a third part with a larger outer diameter, the capillary needle 14 is tubular, and two ends of the capillary needle are open, that is, a through hole penetrates through the first part, the second part and the third part, and the capillary tube is arranged in the through hole; the electrodes are arranged in said capillary 14 with the bulb portion fixed in the third portion and the needle portion extending inside said capillary;
the first clamping jaw is fixed on a base, a gas channel is arranged on the base, an electric conductor is arranged in the gas channel, one side of the base, which is close to the capillary needle 14, is provided with an annular groove surrounding an outlet of the gas channel, and an O-shaped ring is arranged in the annular groove; the base is provided with a first (linear) guide rail, the second clamping jaw is arranged on the second guide rail, and the linear connecting piece is connected with the second clamping jaw and is provided with teeth; the motor is fixed on the base, a gear is fixed on the rotating shaft and meshed with teeth on the connecting piece, and when the motor rotates, the second clamping jaw (connecting piece) is driven to move horizontally on the first guide rail; the first jaw is provided with a first groove, the second jaw is provided with a second groove, when the first jaw and the second jaw are in a vertical static state, the first groove and the second groove respectively comprise an upper wall, a side wall and a lower wall, and the distance from the lower wall to the upper wall is equal to the sum of the heights (in the central axis direction of the capillary needle) of the second part and the third part of the capillary needle; the side wall is divided into a fourth part and a fifth part from bottom to top; from the lower wall to the upper wall, the distance between the side walls of the first groove and the second groove is gradually increased in the fourth portion and is unchanged in the fifth portion, so that when the capillary needle is clamped by the first clamping jaw and the second clamping jaw, the second portion is positioned between the fourth portions of the first clamping jaw and the second clamping jaw, the third portion is positioned between the fifth portions of the first clamping jaw and the second clamping jaw, the O-shaped ring is squeezed by the base and the third portions of the capillary needle, and meanwhile, the electric conductor is in contact with the spherical portion.
The single-cell mass spectrometry method of the embodiment of the invention, namely the working method of the single-cell mass spectrometry system of the embodiment of the invention, comprises the following steps:
(A1) The moving member 11 and the two-dimensional moving platform cooperate to move the carrier 12 to the capillary needle carrier 31 and the holding device 13 to the selected capillary needle 14;
(A2) The driving unit drives the second clamping jaw to move forward on the first guide rail in the horizontal direction, a third part of the capillary needle enters the first groove and/or the second groove firstly, then the second part enters the first groove and/or the second groove, the second part enters the two grooves, the capillary needle moves upwards to enable the capillary needle to be clamped by the first clamping jaw and the second clamping jaw, the second part is located between the fourth parts of the first clamping jaw and the second clamping jaw, the third part is located between the fifth parts of the first clamping jaw and the second clamping jaw, the O-shaped ring is squeezed by the base and the third part of the capillary needle, the gas channel is communicated with the capillary, meanwhile, the electric conductor is contacted with the spherical part, and simultaneous conduction of a circuit and an air circuit is realized;
driving the bearing piece to move upwards, and separating the capillary needle from the bearing device 31;
(A3) The moving member 11 moves forward on the second guide rail 21, and the capillary needle 14 is moved to the single cell carrying device 41 to complete the extraction of the selected single cell, such as generating picoliter extraction liquid and absorbing the extraction material of the single cell, which are the prior art in the field;
(A4) The moving member 11 continues to move forward on the second guide rail 21, the capillary needle 14 is moved to the mass spectrum analyzer 51, the electrodes discharge to realize ionization of the extracted substances, and the ions enter the mass spectrum analyzer and are analyzed to finish the analysis of single cells;
(A5) The driving unit drives the second clamping jaw to move reversely on the first guide rail, the first clamping jaw is separated from the second clamping jaw, and the capillary needle falls and is collected;
the moving member 11 continues to move reversely on the second guide rail to move to the loading position of the capillary needle 14.
Example 3:
the application example of the single-cell mass spectrometry system and method according to embodiment 1 of the present invention is different from embodiment 2 in that:
the first clamping jaw is arranged on the first guide rail, and the first clamping jaw and the second clamping jaw are translated on the first guide rail in opposite directions by using the driving unit.
Claims (10)
1. The single-cell mass spectrometry system comprises a capillary needle, a single-cell bearing device and a mass spectrum analyzer; the capillary needle is characterized by comprising a first part with a smaller outer diameter, a second part with a gradually larger outer diameter and a third part with a larger outer diameter which are distributed in sequence; the single-cell mass spectrometry system further comprises:
the bearing part is arranged on the moving part, the capillary needle is arranged at one end of the bearing part, which is far away from the moving part, and the moving part is used for moving the capillary needle to the single cell bearing device, the mass spectrometer and the capillary needle bearing device respectively;
the capillary needle carrying device is provided with a plurality of carrying positions suitable for carrying the capillary needles;
the clamping device comprises a base, a first clamping jaw, a second clamping jaw, a first guide rail and a driving unit, wherein the base is fixed on the bearing piece, the first clamping jaw is arranged on the base, the first guide rail is arranged on the base, and the driving unit is used for driving the second clamping jaw to move on the first guide rail in the forward direction and the direction; the first clamping jaw and the second clamping jaw are respectively provided with oppositely arranged grooves, and the distance between the side walls of the two grooves is increased from the lower wall of each groove to the upper part of each groove; the second part and the third part are clamped in the two grooves of the first clamping jaw and the second clamping jaw.
2. The single-cell mass spectrometry system of claim 1, further comprising:
the moving piece is arranged on the second guide rail;
the two-dimensional moving platform is arranged on the moving part, and the bearing part is arranged on the two-dimensional moving platform.
3. The single-cell mass spectrometry system of claim 1, wherein the base has a gas channel therein, the capillary needle has a through-hole extending through the first, second and third portions, and the capillary tube is disposed within the through-hole; when the capillary needle is positioned in the two grooves, the through hole is communicated with the gas channel.
4. The single-cell mass spectrometry system of claim 3, further comprising:
an electrical conductor disposed within the gas channel;
the electrode is arranged in the capillary needle; when the capillary needle is in the two grooves, the electric conductor is connected with the electrode.
5. The single-cell mass spectrometry system of claim 4, wherein the electrode comprises a ball portion and a needle portion extending within the capillary; when the capillary needle is in the two grooves, the electric conductor is connected with the spherical part.
6. The single-cell mass spectrometry system of claim 3, wherein the side of the base adjacent the capillary needle has an annular groove surrounding the gas channel opening, and a seal is disposed in the annular groove; when the capillary needle is in the two grooves, the sealing element is pressed by the base and the capillary needle.
7. The single-cell mass spectrometry system of claim 1, wherein the drive unit comprises:
the motor is fixed on the base and drives the gear to rotate;
and the linear connecting piece is arranged on the first guide rail and is connected with the second clamping jaw, and the gear is meshed with the teeth on the connecting piece.
8. The single-cell mass spectrometry method of the single-cell mass spectrometry system of any one of claims 1 to 7, comprising the steps of:
(A1) The moving piece moves the bearing piece to the capillary needle bearing device, and the clamping device moves to the selected capillary needle;
(A2) The driving unit drives the second clamping jaw to move forwards on the first guide rail, and the third part and the second part of the capillary needle enter the two grooves and are clamped by the first clamping jaw and the second clamping jaw.
9. The method of mass spectrometry of a single cell as claimed in claim 8, wherein said method of mass spectrometry of a single cell further comprises the steps of:
(A3) The moving piece moves the capillary needle to the single cell bearing device to complete the extraction of the selected single cell;
(A4) The moving piece moves the capillary needle to the mass spectrometer to complete the analysis of the single cell;
(A5) The driving unit drives the second clamping jaw to move reversely on the first guide rail, the first clamping jaw is separated from the second clamping jaw, and the capillary needle falls off.
10. The method according to claim 8, wherein in step (A2), when the capillary needle is clamped by the first and second clamping jaws, the electrical conductor in the gas channel on the base is connected to the electrode in the capillary needle, and the interior of the capillary needle is in communication with the gas channel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202210370626.6A CN115436451A (en) | 2022-04-10 | 2022-04-10 | Single cell mass spectrometry systems and methods |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210370626.6A CN115436451A (en) | 2022-04-10 | 2022-04-10 | Single cell mass spectrometry systems and methods |
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| CN115436451A true CN115436451A (en) | 2022-12-06 |
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| CN202210370626.6A Pending CN115436451A (en) | 2022-04-10 | 2022-04-10 | Single cell mass spectrometry systems and methods |
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