Open AccessMethods for cell proliferation analysis by fluorescent image cytometry
Author(s) -
Souchier Catherine,
Ffrench Martine,
Benchaib Mehdi,
Catallo Regine,
Bryon Paul Andre
Publication year - 1995
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990200303
Subject(s) - propidium iodide , microbiology and biotechnology , population , fluorescein isothiocyanate , cytometry , flow cytometry , biology , bromodeoxyuridine , dna , cell , fluorescence , chemistry , cell growth , apoptosis , biochemistry , physics , medicine , environmental health , quantum mechanics , programmed cell death
Abstract Methods were developed for multimodal microscopic image analysis in order to identify and analyze one cell type under various microscopic conditions. Our purpose was to quantify both total DNA content using propidium iodide (PI) stain and S‐phase fraction using the bromodeoxyuridine (BrdUrd) incorporation technique in cell population subsets. The model chosen was plasma cells in bone marrow triply labelled with fluorescein isothiocyanate (FITC) for intracytoplasmic immunoglobulins, with amino‐methylcoumarin‐acetate (AMCA) for BrdUrd, and with PI for DNA. Image analysis included three phases. First, plasma cells were recognized on FITC images, and the centroid positions were stored. Second, plasma cell nuclei were geodesically reconstructed from these stored positions using PI images in which DNA content was measured, and the nuclear mask outlines were stored. Third, BrdUrd incorporation level of plasma cells was measured on AMCA images inside PI nuclei masks and stored. Image DNA vs. BrdUrd scatterplots were obtained for cells selected according to the expression of intracytoplasmic immunoglobulin. Thus, both ploidy and proliferation could be independently evaluated on a subset of the cellular population. © 1995 Wiley‐Liss, Inc.