CN215988660U - A high-efficient sampling system for unicellular detection - Google Patents
A high-efficient sampling system for unicellular detection Download PDFInfo
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- CN215988660U CN215988660U CN202122328531.1U CN202122328531U CN215988660U CN 215988660 U CN215988660 U CN 215988660U CN 202122328531 U CN202122328531 U CN 202122328531U CN 215988660 U CN215988660 U CN 215988660U
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- 238000001514 detection method Methods 0.000 title description 15
- 238000005070 sampling Methods 0.000 title description 2
- 239000007789 gas Substances 0.000 description 38
- 239000012159 carrier gas Substances 0.000 description 19
- 239000002245 particle Substances 0.000 description 14
- 238000000889 atomisation Methods 0.000 description 9
- 239000006285 cell suspension Substances 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 8
- 230000035945 sensitivity Effects 0.000 description 8
- 230000009471 action Effects 0.000 description 7
- 230000002093 peripheral effect Effects 0.000 description 7
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 239000000443 aerosol Substances 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 238000005086 pumping Methods 0.000 description 5
- 230000001133 acceleration Effects 0.000 description 4
- 229910052786 argon Inorganic materials 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000012083 mass cytometry Methods 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000000112 cooling gas Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000033001 locomotion Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000004931 aggregating effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- 238000002663 nebulization Methods 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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Abstract
The utility model discloses a high-efficiency sample introduction system for single cell detection, which comprises an atomizer for atomizing a sample to be detected output by an injector to form aerosol, an atomizing chamber for reducing a solvent on the surface of a sample cell with an ultra-large particle size and a concentrator with an inner cavity and an outer cavity which are independent. Based on the aerodynamic principle, the cell sample is concentrated, other cells or impurities outside the cells of the detected sample are removed, the cell number concentration and ionization efficiency entering the rectangular tube are improved, the number of ionization areas of the cell sample in the center of the rectangular tube is increased, and the detection sensitivity of an instrument is improved; at the same sensitivity, the power requirement of the load of the ICP source RF coil is reduced, and the gas consumption for cooling is reduced.
Description
Technical Field
The utility model belongs to the technical field of analysis and detection instruments, and particularly relates to a high-efficiency sample introduction system for single cell detection.
Background
Cells are basic units of organism structure and function, and in order to explore the working mechanism of a living organism, the cells are generally required to be taken as research objects to deeply research the growth, propagation, apoptosis, heredity, evolution and the like of the cells. However, due to the limitation of conditions, the traditional method usually takes the colony cells as the research object, and obtains the information of the cells on the average level. However, there are often great differences between cells, so that it is necessary to establish an analysis method based on single cell level to explain the action mechanism of cells on life activities more accurately. At present, single cell analysis methods based on Mass spectrometry technology mainly include single cell ICP-MS (SC-ICP-MS) and Mass Cytometry (Mass Cytometry), wherein the SC-ICP-MS can directly analyze element types and contents in single cells, and the Mass Cytometry can carry out multi-parameter detection on the single cells. The two technologies are based on ICP plasma source, a single-cell sample to be detected is generated through a front-end sample introduction system, then enters plasma for ionization, and finally is detected through mass spectrum. However, there are two problems: 1) the sensitivity of ICP for detecting cell samples needs to be further improved, and the problem is often solved by improving the radio frequency power of ICP, but the consumption of cooling gas is greatly increased; 2) the sample cell enters the torch tube from the atomizing chamber through the carrier gas, and the whole beam diameter is relatively diverged, so that the ionization efficiency of ICP is influenced.
In view of the above-mentioned problems, the main solution is to improve the detection sensitivity by improving the structure of the torch, the atomizer or the mist chamber. For example, in FIG. 1, Alexander et al, the cell delivery efficiency was 75.0. + -. 4.7% by the newly designed nebulization system. However, more carrier gas and atomizing gas enter the ICP torch, and the number concentration of the sample cells is still lower. The patent (US 2017/0098532) designs a new atomization chamber for cell sample injection, the structure of which is shown in fig. 2, and the atomization chamber is divided into a deceleration area and an acceleration area, and the atomization chamber has a heating function, and can realize desolventization of sample cells. The method belongs to a more traditional method in cell sample introduction, and has limited improvement on detection sensitivity. As shown in FIG. 3, ALAVI et al (WO 2019/144220A 1) have designed a tapered torch which reduces the amount of argon as a cooling gas by 70%, thus increasing the RF power applied to the coil, i.e., increasing the detection sensitivity, under the same conditions. In addition, the researchers also design a recovery system for cooling argon gas to indirectly improve the radio frequency power, but the design is more complex and difficult to popularize.
SUMMERY OF THE UTILITY MODEL
Aiming at the problems in the prior art, the utility model aims to provide a high-efficiency sample introduction system for single cell detection.
The utility model is realized by the following technical scheme:
the high-efficiency sample introduction system for single cell detection is characterized by comprising an atomizer, an atomizing chamber and a concentrator;
the atomizer is connected to the injector through an atomizing capillary tube and atomizes a sample to be detected output by the injector to form aerosol;
the atomizing chamber is provided with an atomizer interface at the front end, a tail end outlet at the rear end, and a carrier gas branch pipe is arranged on the side wall close to the atomizer interface, wherein the atomizer interface is used for inserting and fixing the atomizer along the axial direction of the atomizing chamber so as to introduce aerosol into the atomizing chamber, the carrier gas branch pipe is used for introducing carrier gas into the atomizing chamber, and the carrier gas carries the aerosol to be output from the tail end outlet;
the concentrator, its front end is provided with the atomizer chamber connector for the connection installation of atomizer chamber, the concentrator has inside and outside two independent cavities, and coaxial arrangement has hole and receiving hole with higher speed in the inner chamber, the other end and the quarter bend of receiving hole link to each other, and outer cavity links to each other with peripheral flowmeter and pump through the branch pipe on being close to the lateral wall of exit end.
Further, the atomizer still includes atomizer outer tube and atomizer inner tube with atomizing capillary coaxial arrangement, the atomizer outer tube is close to cell suspension introduction end and is provided with atomizing branch pipe. The atomizer is mainly used for generating a single cell sample, injecting cell suspension through a peripheral sample injection pipeline, injecting the cell suspension through an atomizing capillary, and simultaneously atomizing by adopting atomizing gas.
After the cell suspension is acted by an atomizer, cell samples with different particle sizes are obtained, and meanwhile, atomizing gas also enters an atomizing chamber; in the atomizing chamber, there is generally a heating device for reducing the solvent on the surface of some sample cells with ultra-large particle size; the carrier gas branch pipe in the atomizing chamber is used for introducing carrier gas with a certain flow rate, so that the generated cell sample enters the next-stage concentrator.
In the concentrator, the cell sample is separated from the gas consisting of the carrier gas and the atomizing gas in the accelerating holes. The sample cell has relatively large particle size, generally in the micrometer range, so that the movement inertia of the sample cell is far larger than that of the gas under the action of the accelerating hole. Therefore, a certain flow of gas is pumped by the peripheral flow meter and the pump, so that the flow of the gas entering the next receiving hole is reduced, and the unit volume number concentration of the sample cells is improved.
Furthermore, an extension pipe is arranged on the accelerating hole and extends to the atomizing chamber connecting port for being connected with the tail end outlet of the atomizing chamber.
Further, the aperture of the accelerating hole is smaller than that of the receiving hole, and a gap is left between the accelerating hole and the receiving hole. The aperture of the accelerating hole is optimized to be 0.5mm-1.5mm, and the clearance between the accelerating hole and the receiving hole is optimized to be 1-2 mm.
Furthermore, inner cavity aspirating holes are uniformly distributed on the circumferential wall of the inner cavity, and the inner cavity aspirating holes are arranged corresponding to the gaps between the accelerating holes and the receiving holes. The outer chamber is pumped through the round holes uniformly distributed on the wall of the inner chamber, and the peripheral flow meter and the pump are connected with the side pipe of the outer chamber for pumping.
Furthermore, the accelerating holes are arranged in a conical shape, so that the inertia and gas diffusion of the sample cells can be improved, and the receiving holes are arranged in a reverse conical shape, so that the speed of the received sample cells can be reduced, the contact time between the sample cells and a plasma source can be prolonged, and the ionization efficiency can be improved.
Compared with the prior art, the utility model has the following beneficial effects:
1) by concentrating the cell sample, the cell number concentration entering the rectangular tube is improved, the number of the cell sample in an ionization area at the center of the rectangular tube is increased, and the detection sensitivity of an instrument is improved;
2) through the arranged concentrator, other cells or impurities outside the cells of the detected sample are removed through aerodynamic separation, the ionization efficiency of the sample to be detected is improved, and the detection sensitivity of the system is improved;
3) at the same sensitivity, the power requirement of the load of the ICP source RF coil is reduced, and the gas consumption for cooling is reduced.
Drawings
FIG. 1 is a diagram of a prior art atomization system;
FIG. 2 is a diagram of a prior art cell sample injection chamber;
FIG. 3 is a schematic view of a prior art tapered torch tube configuration;
FIG. 4 is a schematic view of the sample injection system of the present invention;
FIG. 5 is a cross-sectional view of the concentrator of the present invention;
FIG. 6 is a trace diagram of cells of different particle sizes based on CFD software simulation according to the present invention;
FIG. 7 is a flow chart of the present invention as a whole;
in the figure, 1-atomizer, 101-atomization capillary tube, 102-atomizer outer tube, 103-atomizer inner tube, 104-atomization gas branch tube, 2-atomization chamber, 201-atomizer interface, 202-tail end outlet, 203-carrier gas branch tube, 3-concentrator, 301-atomization chamber connecting port, 302-inner chamber, 303-acceleration hole, 304-receiving hole, 305-outer chamber, 306-branch tube, 307-extension tube, 308-gap, 309-inner chamber pumping hole, 4-rectangular tube, 5-peripheral flow meter and 6-pump.
Detailed Description
The present invention will be further described with reference to the accompanying drawings, which are included to illustrate and not limit the utility model.
As shown in FIG. 4, the present invention provides a high efficiency sample injection system for single cell detection, which comprises an atomizer 1, an atomizing chamber 2 and a concentrator 3.
The atomizer 1 is connected to an injector through an atomizing capillary 101, and atomizes a sample to be detected output by the injector to form aerosol; including atomizer outer tube 102, atomizer inner tube 103 and the capillary 101 of atomizing of coaxial setting, atomizer outer tube 102 is close to cell suspension introduction end and is provided with atomizing gas branch pipe 104, and atomizing gas branch pipe 104 is used for letting in atomizing gas, atomizes the cell suspension that awaits measuring, is provided with the flowmeter on it for atomizing gas is let in to the ration.
The front end of the atomizing chamber 2 is provided with an atomizer interface 201, the rear end of the atomizing chamber is provided with a tail end outlet 202, a carrier gas branch pipe 203 is arranged on the side wall close to the atomizer interface 201 and used for introducing a certain amount of carrier gas to enable a generated cell sample to enter the next-stage concentrator 3, and the atomizer interface 201 is used for inserting and fixing the atomizer 1 along the axial direction of the atomizing chamber 2, so that aerosol is introduced into the atomizing chamber 2.
The concentrator 3 is provided with an atomizing chamber connecting port 301 at the front end and used for connecting and installing the atomizing chamber 2, the concentrator 3 is provided with an inner chamber and an outer chamber, an accelerating hole 303 with a conical structure and a receiving hole 304 with a reverse conical structure are coaxially installed in the inner chamber 302, an extension pipe 307 is arranged on the accelerating hole 303 and extends to the atomizing chamber connecting port 301 to be connected with a tail end outlet of the atomizing chamber 2, the other end of the receiving hole 304 is connected with a rectangular pipe 4, and the side wall of the outer chamber 305 close to the outlet end is connected with a peripheral flow meter 5 and a pump 6 through a branch pipe 306. The aperture of the accelerating holes 303 is smaller than that of the receiving holes 304, gaps 308 are reserved between the accelerating holes 303 and the receiving holes 304, and inner cavity suction holes 309 are uniformly distributed on the circumferential wall of the inner cavity 302 at annular positions corresponding to the gaps 308.
As shown in fig. 5, a half-sectional view of the concentrator 3, it can be seen that the left side of the structure is an acceleration hole 303 connected to the atomization chamber 2 through an extension pipe 307. The aperture of the accelerating hole 303 is 0.8mm, the aperture of the receiving hole 304 is 1mm, and the end faces of the two holes are 1mm apart. In the structure of the concentrator 3, six inner cavity pumping holes 309 are uniformly distributed on the wall of the inner cavity 302, the outlet side of the outer cavity 305 is provided with a branch pipe 306 for connecting the peripheral flow meter 5 and the pump 6. The outlet of the concentrator 3 is connected with a torch pipe 4.
The use method of the sample introduction system comprises the following specific steps:
1) introducing the prepared cell suspension to be detected into the atomizer 1 through a peristaltic pump or an injection pump or other modes;
2) atomizing gas is introduced through the atomizing gas branch pipe 104 by a flowmeter, the flow rate of the atomizing gas is generally low for cell samples, and the flow rate of the atomizing gas is generally 0.15-0.35L/min;
3) under the action of atomizing gas, the cell suspension is atomized by an atomizer 1 and enters an atomizing chamber 2, the particle size of the cells of a sample is generally below 10 mu m, the central particle size is several microns, and cells with the particle size larger than 10 mu m and cells with smaller particle size are also present;
4) the carrier gas is introduced into the atomizing chamber 2 through the carrier gas branch pipe 203 by the flowmeter, the cell sample is brought into the next-stage system, and the general flow is about 1L/min. The atomizing chamber 2 can reduce the solvent on the surface of the cell sample by heating, reduce the particle size of the cells, and simultaneously prevent the cell sample from aggregating into large clusters on the surface of the atomizing chamber 2; the heating device arranged on the atomizing chamber 2 is the prior art, and is not described in detail, and the prior art is referred to;
5) the cell sample enters the concentrator 3 by the action of the atomizing gas and the carrier gas, and since the gas (or small-particle-size cell sample) has a much smaller inertia than the cell sample (of the order of μm), the gas expands and diverges at the acceleration hole, and the cell sample can rapidly pass through the acceleration hole 303 and then enter the receiving hole 304. In the process of gas-particle separation, an external pump and a flowmeter are adopted to cooperate to extract gas, and finally the gas is discharged through a peripheral waste discharge system. For example, the total flow rate of the atomizing gas and the carrier gas is 1L/min, and the pump pumps the atomized gas and the carrier gas at 0.9L/min, so that the flow rate of the residual gas entering the torch tube along with the cell sample is 0.1L/min, namely, the cell number concentration is increased by 10 times, which is expressed on the mass spectrum signal, and the signal to be measured is theoretically increased by 10 times;
6) cell sample is separated by gas particles, so that the concentration of the cell of the sample is concentrated, and the cell enters the torch tube 4 under the action of residual gas. Meanwhile, based on the method, the method has the additional advantages that the amount of the atomizing gas and the carrier gas entering the ICP source is greatly reduced, the influence of the gas on the distribution of the plasma center torch can be effectively reduced, and the small-particle-size cell sample is pumped out along with the pump due to weak inertia, so that the consumption of the small-particle-size cell sample on the ICP can be reduced. On the other hand, considering the problem of consumption of cooling argon, we can reduce the cooling argon under the condition of properly reducing the load power of the RF coil, and simultaneously keep the detection sensitivity of the instrument at a relatively good level.
Based on the established simple concentrator model, the sample cell movement trajectory simulation was performed on the structure by CFD software, as shown in fig. 6. Selecting cells with three particle diameters of Dp =0.1 μm, 0.5 μm, 2 μm and the like as research objects, enabling the total air inlet flow to be 1L/min, enabling the cells to enter a concentrator along with sample cells, enabling the pump pumping flow to be 0.1L/min, and coupling a CFD module and a particle tracking module to obtain the motion trail of the sample cells. It was found that when Dp =0.1 μm, the fraction of sample cells passing through the receiving well was only 8%, mostly excluded with the pump; when Dp =0.5 μm, the proportion of sample cells entering the receiving well with inertia is 42%; when Dp =2 μm, the sample cells can achieve 100% extraction, and the overall beam diameter is restricted. Therefore, based on this method, efficient extraction of sample cells can be achieved.
The process of the utility model is shown in figure 7, the cell suspension is introduced by a syringe pump or an automatic sample injector, and is input into the atomizer through a hose, the sample is output by the atomizing capillary in the middle of the atomizer, and is atomized under the action of the branch atomizing gas. The single cell sample enters an atomizing chamber, and a heating device is wrapped around the atomizing chamber and used for removing the solvent of the sample cells with the ultra-large particle size. Under the action of the carrier gas and the atomizing gas, the sample cells enter the accelerating holes. At the moment, because the periphery is provided with the flowmeter and the pump, the separation of the sample cells and the gas is realized by pumping a certain gas flow, namely the sample cells are concentrated and then pass through the receiving hole, the moving speed of the sample cells is reduced at the receiving hole, then the sample cells enter the torch tube along with the residual gas, and the desolventizing, the gasifying and the ionizing are realized at the plasma source at the tail end of the torch tube. And finally, enabling the ions to pass through a vacuum interface to enter mass spectrum detection.
Claims (7)
1. A high efficiency sample introduction system for single cell detection, the system comprising:
the atomizer (1) is connected to the injector through an atomizing capillary (101) and atomizes a sample to be detected output by the injector to form aerosol;
the atomizing device comprises an atomizing chamber (2), wherein the front end of the atomizing chamber is provided with an atomizer interface (201), the rear end of the atomizing chamber is provided with a tail end outlet (202), a carrier gas branch pipe (203) is arranged on the side wall close to the atomizer interface (201), the atomizer interface (201) is used for inserting and fixing an atomizer (1) along the axial direction of the atomizing chamber (2), so that aerosol is introduced into the atomizing chamber (2), the carrier gas branch pipe (203) is used for introducing carrier gas into the atomizing chamber (2), and the carrier gas carries the aerosol and is output from the tail end outlet (202);
concentrator (3), its front end is provided with atomizer chamber connector (301) for the connection installation of atomizer chamber (2), concentrator (3) have inside and outside two independent cavities, and coaxial arrangement has hole (303) and receiving hole (304) with higher speed in inner chamber (302), the other end and rectangular pipe (4) of receiving hole (304) link to each other, and outer cavity (305) link to each other with peripheral flowmeter (5) and pump (6) through branch pipe (306) on being close to the lateral wall of exit end.
2. The high-efficiency sample introduction system for single cell detection as claimed in claim 1, wherein the atomizer (1) further comprises an outer atomizer tube (102) and an inner atomizer tube (103) coaxially mounted with the atomizing capillary (101), and the outer atomizer tube (102) is provided with an atomizing air branch tube (104) near the sample introduction end of the cell suspension.
3. The high-efficiency sample introduction system for single cell detection as claimed in claim 1, wherein the accelerating hole (303) is provided with an extension tube (307) extending to the connection port of the atomizing chamber (2) for connecting with the tail end outlet of the atomizing chamber (2).
4. A high efficiency sample introduction system for single cell detection as claimed in claim 1, wherein the aperture of said acceleration hole (303) is smaller than the aperture of said receiving hole (304), and a gap (308) is left between said acceleration hole (303) and said receiving hole (304).
5. A high efficiency sample introduction system for single cell detection as claimed in claim 4, wherein the aperture of said acceleration and receiving holes is between 0.5mm and 1.5mm, and said gap is in the range of 1-2 mm.
6. The high-efficiency sample introduction system for single cell detection as claimed in claim 1, wherein inner cavity air extraction holes (309) are uniformly distributed on the circumferential wall of the inner cavity (302), and the inner cavity air extraction holes (309) are arranged corresponding to the gap (308) between the acceleration hole (303) and the receiving hole (304).
7. The high efficiency sample introduction system for single cell detection as claimed in claim 1, wherein the acceleration hole (303) is tapered, and the receiving hole (304) is reversely tapered.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113782413A (en) * | 2021-09-26 | 2021-12-10 | 谱视科技(杭州)有限公司 | Efficient sample introduction system for single cell detection and use method |
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- 2021-09-26 CN CN202122328531.1U patent/CN215988660U/en not_active Withdrawn - After Issue
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113782413A (en) * | 2021-09-26 | 2021-12-10 | 谱视科技(杭州)有限公司 | Efficient sample introduction system for single cell detection and use method |
| CN113782413B (en) * | 2021-09-26 | 2024-10-18 | 谱视科技(杭州)有限公司 | Efficient sample injection system for single cell detection and use method |
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