Fluorochrome choices for multi-color flow cytometry.

Fluorochrome choices for multi-color flow cytometry.
J Immunol Methods. 2019 Jun 07;:
Authors: Flores-Montero J, Kalina T, Corral-Mateos A, Sanoja-Flores L, Pérez-Andrés M, Martin-Ayuso M, Sedek L, Rejlova K, Mayado A, Fernández P, van der Velden V, Bottcher S, van Dongen JJM, Orfao A
Abstract
Fluorochrome selection is a key step in designing multi-color antibody panels. The list of available fluorochromes is continuously growing, fitting current needs in clinical flow cytometry to simultaneously use more markers to better define multiple leukocyte subpopulations in a single tube. Several criteria guide fluorochrome selection: i) the fluorescence profiles (excitation and emission), ii) relative brightness, iii) fluorescence overlap, iv) fluorochrome stability, and v) reproducible conjugation to antibodies. Here we used 75 samples (45 bone marrow and 30 blood) to illustrate EuroFlow strategies for evaluation of compatible fluorochromes, and how the results obtained guide fluorochrome selection as a critical step in the antibody-panel building process. Our results allowed identification of optimal fluorescence profiles (e.g. higher fluorescence intensity and/or resolution with limited fluorescence overlap into neighbor channels) for brilliant violet (BV)421 and BV510 in the violet laser and allophycocyanin (APC) hilite 7 (H7) or APC C750 in the red laser vs. other candidate fluorochromes generally applied for the same detectors and ...

Source: Journal of Immunological Methods - June 6, 2019 Category: Allergy & Immunology Authors: Flores-Montero J, Kalina T, Corral-Mateos A, Sanoja-Flores L, Pérez-Andrés M, Martin-Ayuso M, Sedek L, Rejlova K, Mayado A, Fernández P, van der Velden V, Bottcher S, van Dongen JJM, Orfao A Tags: J Immunol Methods Source Type: research