Identification method of nucleated cells and application thereof

Abstract

The invention provides an identification method of nucleated cells and an application thereof, the identification method of the nucleated cells can be applied in a single-cell separation detection platform with the combination of the micromanipulator capture method and LV-PCR and the single-cell separation detection platform with the combination of the laser microdissection method and the LV-PCR,and the method comprises the following steps: placing a sample containing the nucleated cells in a centrifugal tube containing buffer solution, oscillating the centrifugal tube to lead the cells on the sample to fall off, and obtaining cell suspension of the cells of the sample; and adding gentian violet or haematoxylin into the cell suspension for dyeing, wherein the final concentration of the gentian violet is 0.1-0.5 mu g / mu l, the dyeing time is 2-7 minutes, the final concentration of the haematoxylin is 1-7 mu g / mu l, and the dyeing time is 3-7 minutes. The use of the cell identificationmethod in the two single-cell separation detection platforms can realize effective identification of the cells of the sample, lead the follow-up LV-PCR amplification inhibition to be weak and obtain good DNA typing result.

Description

technical field [0001] The invention relates to the fields of biotechnology and forensic science, in particular to a nucleated cell marking method suitable for a single cell separation and detection platform and its application. Background technique [0002] In the detection of criminal cases, many of them require the intervention of forensic identification. In most cases, the specimens obtained on site are usually difficult and mixed specimens. Compared with the ideal specimens for research, the prominent problem is that the number of target cells in the specimens obtained is small and the morphology changes greatly. The boundary between nucleus and cytoplasm is unclear, and the background is complex. In the absence of cell labeling, the target cells on these specimens cannot be found or are easily misidentified, resulting in the inability to obtain the definite DNA typing results of the specimen cells. Therefore, specific and effective cells must be used for these specime...

AI Technical Summary

Problems solved by technology

[0004] There have also been some studies on the isolation of spermatids from simulated mixed seminal spot samples using laser capture microdissection (LCM), using five histological staining methods (HE, CTS, AO, Wright's, methyl Green method) to stain the smears of sperm cells and vaginal epithelial cells, and compare the labeling effect and the degree of inhibition of PCR amplification. PCR amplification inhibition

[0005] Although the above-mentioned staining method can basically satisfy the detection of ideal specimens, for the micro-mixed specimens from accidents or cases, because the dyes significantly inhibit the PCR amplification, even if the above staining method is used to obtain a good cell labeling effect, Final genotyping results are still highly biased

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