Open AccessLight scatter analysis and sorting of cells activated in mixed leukocyte culture
Author(s) -
MacDonald H. Robson,
Zaech Pierre
Publication year - 1982
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990030112
Subject(s) - sorting , cytometry , flow cytometry , microbiology and biotechnology , computational biology , biology , computer science , algorithm
Cells activated in unidirectional mixed leukocyte cultures (MLC) have been analyzed on the basis of their light scatter characteristics. C57BL/6 spleen cells were cultured with irradiated (2000 rads) DBA/2 spleen cells for 5 days and the resulting suspension of activated cells was passed on a FACS II flow cytometer. Correlated parameter analysis of forward light scatter (FLS) and perpendicular light scatter (PLS) indicated that the MLC consisted of a heterogenous mixture of viable cells, dead cells, and subcellular debris. However, by appropriate gating of the FLS/PLS distribution, viable cells could be identified as a biphasic FLS histogram. Sorting and morphological analyses of these two FLS peaks demonstrated that they corresponded to almost pure populations of small lymphocytes (lower peak) and lymphoblasts (upper peak), respectively. Furthermore, when sorted cells were tested for their ability to lyse antigenically relevant (DBA/2) tumor target cells in a 51 Crrelease assay, lymphoblasts were found to exhibit 40‐fold greater cytolytic activity (on a per‐cell basis) than small lymphocytes.