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Rapid cell classification and quantification method based on coffee ring

A technology of coffee rings and cells, which is applied in the field of cell detection, can solve the problems of high sample carrier cost and low signal-to-noise ratio, and achieve the effects of quantitative accuracy, improved signal-to-noise ratio, and high linear correlation

Pending Publication Date: 2021-07-09
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to overcome the influence of the coffee ring effect, CN 110927143 A discloses that the sample detected by LIBS is enriched in the annular groove, which can greatly reduce the uneven enrichment problem caused by the coffee ring effect, but the cost of such a sample carrier is relatively high
Some detections avoid the coffee ring area during sampling, and only select the relatively uniform area in the middle but with a sparse number of cells, but the signal-to-noise ratio of this is very low

Method used

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  • Rapid cell classification and quantification method based on coffee ring
  • Rapid cell classification and quantification method based on coffee ring
  • Rapid cell classification and quantification method based on coffee ring

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1 Rapid cell detection method of the present invention

[0061] The rapid cell detection method of the present invention comprises the steps of:

[0062] 1. Preparation of Silicon Wafer Substrate

[0063] The 8×8mm monocrystalline silicon wafer was ultrasonically cleaned with acetone, ethanol and ultrapure water for 10 minutes, then soaked in piranha solution (ie piranha lotion) for 10 minutes, and then washed with ultrapure water for 3 times. Afterwards, the silicon wafer was soaked in hydrofluoric acid (5% by volume) for 3 minutes to complete the treatment of the substrate. At this time, the surface of the silicon wafer was clean and had a certain degree of hydrophobicity.

[0064] 2. Preparation of Cell Suspension

[0065] The cells cultured to the logarithmic phase were digested with trypsin to obtain a cell suspension, centrifuged at 1200r / min for 3 minutes to obtain a cell pellet, and the cells were washed twice with buffer and then resuspended to eli...

experiment example 1

[0074] Rapid Classification of Several Cells in Experimental Example 1

[0075] 1. Method

[0076] Human normal somatic cells and cancer cells are distinguished respectively. The normal human somatic cells are the WRL-68 human liver cell line, and the cancer cells include the Huh-7 liver cancer cell line and the A549 human non-small cell lung cancer cell line. In the same (10 6 cells / mL), 4 μL of the cell suspension was dropped onto the silicon chip analysis substrate, and after natural drying, a unique fingerprint of the distribution of cells on the substrate was obtained (the process of drying the sample to form a coffee ring is as follows: Figure 4 As shown, the fingerprints are as Figure 5 shown). The silicon substrate was fixed on the stage of the LIBS detection instrument for further analysis. The conditions of laser energy, accumulation times and delay time parameters were optimized respectively. Spectrum collection was performed under the optimal test conditions...

experiment example 2

[0081] Experimental example 2 The linear relationship between the cell concentration and the endogenous characteristic element content of the cells on the coffee ring

[0082] 1. Method

[0083] Take the A549 human non-small cell lung cancer cells of logarithmic growth, and obtain a concentration of 10 3 -10 7 cells / mL cell suspension. Take 4 μL of the cell suspension and drop it on the silicon chip analysis substrate, dry it naturally, fix it on the circular rotary displacement stage, scan the coffee ring area along the circular path, collect the spectral line signals of the endogenous characteristic elements of the cell, and accumulate the endogenous cell The spectral line intensity value of the characteristic element. The conditions of laser energy, accumulation times and delay time parameters were optimized respectively. Under the optimal test conditions of 40% energy, 100 accumulations, and 1.5μs delay time, spectral line collection and accumulation are performed. Th...

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Abstract

The invention discloses a rapid cell classification and quantification method based on a coffee ring and belongs to the field of cell detection. The key point of the invention is to provide an LIBS signal acquisition method of a cell suspension coffee ring, which mainly comprises the following steps of dropwise adding a cell suspension to a monocrystalline silicon wafer, naturally drying to form a coffee ring, and then carrying out LIBS signal acquisition on endogenous characteristic elements of cells in a coffee ring area. The invention further provides a method for constructing the cell characteristic annular imprinting fingerprint element atlas database and the corresponding cell characteristic annular imprinting fingerprint element atlas database. The invention also provides a cell classification method which is used for realizing cell classification by comparing the sample annular mark fingerprint element atlas with the atlas database. The invention also provides a cell quantification method based on the LIBS signal acquisition method of the cell suspension coffee ring. The method realizes quantification based on the linear relationship between a spectral line intensity value and the cell concentration in the LIBS signal.

Description

technical field [0001] The invention belongs to the field of cell detection. Background technique [0002] The classification and quantification of cells are widely used in modern medical testing. [0003] Cell classification methods mainly include staining microscopy, flow cytometry, and VCS combined detection technology. Among them, the staining microscopy method is to use the affinity of cells to acid-base dyes or the activity of intracellular peroxidase to stain with specific dyes and then distinguish them under the microscope. Eosinophils, neutrophils, monocytes) are useful and cannot classify the vast majority of cells. Flow cytometry needs to label the cells with fluorescent antibodies, and the cells with specific fluorescence can be detected by flow cytometry, which cannot detect trace samples, and the cost of reagents is high. The VCS joint detection technology uses electrical impedance to detect cell volume, conductance radio frequency method to detect cell nucl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N15/10
CPCG01N21/31G01N15/10G01N2015/1006
Inventor 林庆宇殷鹏鲲康梦珂段忆翔
Owner SICHUAN UNIV
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