Centrifugal layering enrichment method for tumor cells in bloody body fluid sample

Abstract

The invention relates to a centrifugal layering enrichment method for tumor cells of a bloody body fluid sample. The method specifically comprises the following steps: collecting cases with lung adenocarcinoma primary lesions and hydrothorax metastasis, and reserving an unfixed fresh bloody body fluid sample; removing macroscopic proteins and blood clots in the sample, shaking up the sample, centrifuging, and observing the layering situation of the supernatant and the precipitate; observing the size and the movement condition of the precipitate of the sample with the layering; when layering occurs, taking all tube bottom precipitates when the precipitates are less than 2 mL, and taking blood sedimentation surface matters when the precipitates are more than 2 mL and are fixed at the tube bottom and do not move; when layering occurs, taking movable precipitates, uniformly mixing the precipitates with a red cell breaking liquid according to a volume ratio of 1:(1-4), breaking red cells, and taking red cell breaking precipitates; and when layering does not occur, selecting a sample which cannot be layered, uniformly mixing the sample with the red cell breaking liquid according to a volume ratio of 1:(1-4), breaking red cells, and taking out a red cell breaking precipitate.

Description

technical field [0001] The invention relates to a centrifugal layered enrichment method for tumor cells in bloody body fluid samples, and belongs to the technical field of cytopathological analysis in medical pathology. Background technique [0002] Serosa cavity spread may occur in almost all malignant tumors of the human body system, most commonly seen in lung cancer, gastrointestinal cancer, breast cancer, ovarian cancer, etc. Among them, the metastasis of lung adenocarcinoma is the most common, mostly caused by tumor cell destruction or tumor thrombus obstruction It is caused by lymphatic vessels and blood vessels, so bloody serous effusion is often formed, and early diagnosis can often be made through cytological detection of serous effusion. The bloody pleural effusion or pericardial effusion produced by the metastasis of thymic carcinoma to the thymus or pericardium can appear red in different shades to the naked eye. For the deep red thick blood pleural effusion, the...

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Problems solved by technology

[0003] At present, % acetic acid ethanol is used in the cytological detection of serous cavity effusion to deal with bloody effusion. The expansion of tumor cells by acetic acid and the contraction of tumor cells by ethanol can offset each other. The background of erythrocyte lysis in pleural effusion is clean. However, the fragments of red blood cells dissolved by acetic acid in bloody pleural effusion that cannot be stratified during this process will deepen the background of staining. In addition, it was found in the experiment that the bloody pleural effusion of metastatic lung adenocarcinoma, alone In cases where acetic acid or a mixed solution was used as a redness-breaking treatment agent, TTF-1 antibody nuclear markers were absent or less expressed, which may be due to the rapid precipitation of nuclear proteins by acetic acid and the loss of TTF-1 protein

In the prior art, red blood cells are also dissolved with Xinpai’s cell cleaning solution. This type of cleaning solution is mainly composed of alcohols. For bloody pleural effusions whose sediment will move after centrifugation and stratification, the effect of enriching tumor cells is good. The class will precipitate invisible proteins in bloody pleural effusion that cannot be stratified, causing tumor cells to disperse and cells to shrink slightly

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