Identification of eosinophils in lysed whole blood using side scatter and CD16 negativity

Abstract

The identification of eosinophils in lysed whole blood by flow cytometry can be problematic, since these cells overlap significantly with the neutrophil cluster on forward scatter versus side scatter plots of whole blood samples. Current methods can be time‐consuming when running multiple samples or may compromise yield in the interests of greater accuracy. The use of eosinophil purification techniques prior to FACS analysis or sorting as a way of ensuring purity may have unpredictable effects on eosinophil activation, leading to questionable data interpretation. Here we describe a simple, single‐step method for definition of eosinophils utilizing their high side scatter and CD16 fluorescence negativity to differentiate them from neutrophils. The purity of the neutrophil and eosinophil populations sorted with this gate is close to 100% regardless of the peripheral blood eosinophil count, while the population obtained by sorting on a plot of FSC/SSC was a mixture of eosinophils and neutrophils. We suggest this method as a simple, reproducible, and accurate way of defining eosinophils by flow cytometry for analysis or sorting. Cytometry 30:313–316, 1997. © 1997 Wiley‐Liss, Inc.

The pattern of GPI-80 expression is a useful marker for unusual myeloid maturation in peripheral blood

“…CD15, CD16 and CD33 are often used for the separation of myeloid populations [36,37]. GPI-80 expression was compared to expression of other markers (Supporting information, Fig.…”

Expression of membrane complement regulators, CD46, CD55 and CD59, in mesothelial cells of patients on peritoneal dialysis therapy

“…The neutrophil populations were obtained by gating based on a plot of FSC/SSC versus a plot of SSC/CD16 (Gopinath and Nutman, 1997). The mean fluorescence intensity (MFI) of CReg was evaluated under FACS analysis on a BD FACS-Canto TM II (Becton Dickinson, San Jose, CA).…”

Elevation of Conjunctival Epithelial CD45INTCD11b+CD16+CD14−Neutrophils in Ocular Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis

“…This gating strategy discriminates neutrophils from other granulocytes (such as basophils and eosinophils) and monocytes. 17,18 A significant increase in the number and percentage of neutrophils (186 vs. 3.4, P < 0.01;19% vs. 0.5%, P ¼ 0.001) was seen in SJS-TEN (Fig. 2C).…”