CTC identification and characterization in prostate cancer using K/V protocol and imaging flow cytometry - part 7

This project aimed to identify and characterize circulating tumor cells (CTCs) in peripheral blood of 33 prostate cancer patients. Fraction of peripheral blood mononuclear cells putatively containing CTCs was isolated using density gradient centrifugation. Pan-keratin/vimentin/CD45/CD31/DAPI immunofluorescent staining was performer and DAPI+ or DAPI+CD45/CD31- objects were collected using Amnis® ImageStream®XMk II, Cytek® Biosciences, Inc (lasers: 405 nm, 488 nm, 642 nm, magnification 40x, slow speed flow).

本研究旨在对33例前列腺癌患者外周血中的循环肿瘤细胞（circulating tumor cells, CTCs）开展鉴定与特征分析。通过密度梯度离心法分离得到疑似含有循环肿瘤细胞的外周血单个核细胞组分。完成泛角蛋白/波形蛋白/CD45/CD31/DAPI免疫荧光染色后，使用Cytek® Biosciences, Inc出品的Amnis® ImageStream®XMk II成像流式细胞仪收集DAPI阳性或DAPI阳性且CD45/CD31阴性的目标细胞；该仪器配备405 nm、488 nm、642 nm激光器，放大倍数为40倍，采用低速流模式。