METHOD FOR DETECTING NEUTROPHIL CHEMOTAXIS

摘要

A method for detecting neutrophil chemotaxis, comprising: collecting a blood sample into an anticoagulant tube, adding glucose, uniformly mixing, and standing; taking the supernatant into a centrifuge tube, and centrifuging same; adding 1× calcium and magnesium ion-free Hanks balanced salt solution into the centrifuged blood sample to re-suspend cell aggregates at the bottom; uniformly blowing and dispersing same, and slowly adding a polysucrose solution from the bottom of the centrifuge tube to perform density gradient centrifugation treatment; the solution after density gradient centrifugation being divided into three layers, pipetting the clear liquid at the upper layer and the PBMC layer at the middle layer to obtain cell aggregates at the bottom layer; performing lysis treatment on erythrocytes in the cell aggregates by using sterile water, performing gentle blowing and suction, then successively adding 2× calcium and magnesium ion-free Hanks balanced salt solution and 1× calcium and magnesium ion-free Hanks balanced salt solution, uniformly mixing same, and performing centrifugation treatment again to obtain neutrophils; and counting the neutrophils, and pipetting the neutrophil suspension into a chemotaxis model for chemotaxis. The method can improve the speed of neutrophil extraction and shorten the detection time.