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Oil-free sorting-direct injection-ICPMS single cell analysis system

An analysis system and single-cell technology, applied in the field of single-cell analysis platform, can solve the problems of limiting the throughput of cell analysis, limiting the efficiency of cell detection, and failing to meet high throughput and high efficiency

Active Publication Date: 2020-06-12
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in order to ensure that two or more cells will not be wrapped in one droplet, the concentration of the cell suspension needs to be reduced, which limits the throughput of cell analysis; the use of the oil phase seriously affects the ionization efficiency of ICPMS and the mass spectrometer itself Stability; and the interface between the existing link cell sorting chip and ICPMS, because it is equipped with a larger volume spray chamber, the transmission efficiency (sample injection efficiency) of the cells is very low, which limits the improvement of the cell detection efficiency
In conclusion, the existing technology cannot satisfy high-throughput and high-efficiency single-cell ICPMS analysis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Determination of Zn in HeLa cells

[0043] 1. Double "straight-bend-straight" microfluidic chip. The preparation of the chip is as described above, and the dimensions and specifications of the microfluidic chip used in this example are as follows: the height of the channels used is 46 μm, where the NH 4 HCO 3 The inlet 1 of the buffer conditioner and the inlet 3 of the suspension mainly consist of a cylindrical structure with a radius of 550 μm, which is used for sample injection, and the channel at the rear end of the inlet is tapered to 100 μm, and is connected with the channel for cell separation at the back. There are also 56 hexagonal columns with a diameter of 50 μm, which are used to filter impurities in samples and pre-separate cells. The first-stage cell separation channel 4 consists of 12 straight-curved structures (12×1500 μm long×100 μm wide straight channels and 14 curved channels with an outer diameter of 150 μm and an inner diameter of 50 μm)...

Embodiment 2

[0052] Example 2: Determination of Au in HeLa cells incubated with AuNPs

[0053] 1. Double "straight-bend-straight" microfluidic chip. The size specification and preparation method of the chip are the same as those in Example 1.

[0054] 2. Install the direct injection micro-spray device. Its dimension specification and installation method are identical with embodiment 1.

[0055] 3. Treatment of cells. Add gold nanoparticles with a size of 15 nm to a final concentration of 10 μg / mL to a plate of growing HeLa cells at 37 °C, 5% CO 2 Cultivate for 4 hours under the same conditions. Afterwards, the cells were enzymatically digested from the bottom of the culture dish with trypsin, and washed 5 times with 1×PBS to remove the medium adsorbed on the cell surface. After the last wash, the cells were pelleted by centrifugation and resuspended in 10 mmol / L NH 4 HCO 3 buffer, perform a cell count. Dilute the cell suspension with NH 4 HCO 3 Buffer diluted to 1.8 x 10 4 cells...

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Abstract

The invention discloses an oil-free sorting-direct injection-ICPMS single cell analysis system, and relates to a single cell analysis platform. A sampling device integrating cell sorting, control andsingle cell direct injection is coupled with ICPMS mass spectrometry detection to perform efficient single cell analysis. The system comprises a double-straight-line-bending-straight-line type micro-fluidic chip body and a direct injection type micro-spraying device. The double-straight-line-bending-straight-line type micro-fluidic chip comprises a buffer solution inlet, a buffer solution channel,a suspension liquid inlet and a two-stage separation channel; a buffer solution and suspension liquid inlet main body is of a cylindrical structure, and a three-fork type connected two-stage separation channel is composed of a plurality of linear-bent-linear structures; the direct injection type micro-spraying device comprises a shell with a carrier gas branch pipe, and the tail end of the shellis gradually thinned to form a nozzle; the sample injection capillary tube is inserted from the tail end of the shell and slightly retracts relative to the nozzle, the polyimide coating on the surfaceof one section of capillary tube is stripped, and a micro cavity is formed on line at the nozzle. An ammonia bicarbonate buffer solution is used in the whole system.

Description

technical field [0001] The invention relates to a single-cell analysis platform, in particular to an oil-free sorting-direct injection-inductively coupled plasma mass spectrometry (ICPMS) single-cell analysis system. Background technique [0002] With the continuous development of life sciences and precision medicine, the importance of single-cell analysis has become increasingly prominent. Microfluidic technology is an ideal single-cell manipulation technology (1. Di Carlo, D.; Irimia, D.; Tompkins, R.G.; Toner, M. Continuous Inertial Focusing, Ordering, and Separation of Particles in Microchannels. P. Natl.Acad.Sci.USA2007,104,18892–18897), while inductively coupled plasma mass spectrometry (ICPMS) has extremely high sensitivity and isotope dilution-multi-element simultaneous quantitative detection ability (2.Inductively Coupled Plasma Mass Spectrometry and Its Applications. 2nded. Edited by Steve J. Hill. Blackwell Pub., 2006). The organic combination of the two facilit...

Claims

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Application Information

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IPC IPC(8): B01L3/00G01N27/62
CPCB01L3/502707B01L3/502753G01N27/62
Inventor 王秋泉周阳陈张倩
Owner XIAMEN UNIV
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