用于联合测量白色念珠菌吞噬作用和细胞内杀伤的流式细胞术检测

Flow cytometric assay for combined measurement of phagocytosis and intracellular killing of Candida albicans.

作者信息

Bjerknes R

出版信息

J Immunol Methods. 1984 Aug 3;72(1):229-41. doi: 10.1016/0022-1759(84)90451-4.

Abstract

A rapid quantitative flow cytometric (FCM) assay for the combined kinetic measurement of phagocytosis and intracellular killing of fluorescein-isothiocyanate (FITC) labelled Candida albicans is described. The fraction of phagocytosing leukocytes and the numbers of attached or internalized Candida albicans per phagocytosing leukocyte were quantified by FCM, using trypan blue and a fluorescence quenching technique. Results obtained by microscopy agreed with the FCM estimates of phagocytosis. Dead, but not live, Candida albicans stained by propidium iodide (PI). Thus, both viable and intracellularly killed fungi could be discriminated and measured by FCM. Phagocyte killing determined by the FCM assay correlated with killing measured by a standard microbiological test and by methylene blue staining. The method allows rapid and accurate testing of opsonic and phagocytic functions under both experimental and clinical conditions.

摘要

本文描述了一种快速定量流式细胞术（FCM）检测方法，用于联合动力学测量吞噬作用以及对异硫氰酸荧光素（FITC）标记的白色念珠菌的细胞内杀伤。使用台盼蓝和荧光猝灭技术，通过FCM对吞噬白细胞的比例以及每个吞噬白细胞上附着或内化的白色念珠菌数量进行定量。显微镜检查获得的结果与FCM对吞噬作用的估计结果一致。碘化丙啶（PI）可对死的而非活的白色念珠菌进行染色。因此，通过FCM可以区分和测量存活的以及细胞内被杀伤的真菌。FCM检测所确定的吞噬细胞杀伤作用与标准微生物学检测及亚甲蓝染色所测量的杀伤作用相关。该方法能够在实验和临床条件下快速、准确地检测调理素和吞噬功能。