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Bio‐maleimide as a generic stain for detection and quantitation of microparticles
Author(s) -
ENJETI A. K.,
LINCZ L.,
SELDON M.
Publication year - 2008
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/j.1751-553x.2007.00937.x
Subject(s) - stain , maleimide , staining , flow cytometry , chemistry , annexin , microbiology and biotechnology , cytometry , incubation , chromatography , pathology , medicine , biochemistry , biology , polymer chemistry
Summary Microparticles (MP) are small fragments of cytoplasm shed from a cell surface and their role in the pathophysiology of disease is being extensively investigated. A novel staining technique for quantifying total MP in peripheral blood was evaluated in this study. Evaluation of Bodipy‐maleimide (or bio‐maleimide) as a stain for quantifying total MP in peripheral blood by flow cytometry. Samples were obtained from 10 healthy donors after informed consent. Plasma was prepared by sequential centrifugation at 1500 g followed by 13 000 g and stained with Annexin V and bio‐maleimide. Enumeration beads were added after 15 min of incubation with the stain and samples analyzed on a FACS Canto flow cytometer. Detection and quantification of MP by bio‐maleimide staining was comparable with that by Annexin V. The total mean MP level with bio‐maleimide staining was 34 ± 19.7/ μ l (range of 11.6–68.1/ μ l) and with Annexin V staining it was 38.9 ± 29.8/ μ l (range of 10.6 to 112.9/ μ l). There was no significant difference using a paired t ‐test and methods were comparable using a Bland–Altman plot. Bio‐maleimide is a useful and inexpensive stain to measure total MP levels in peripheral blood by flow cytometry. This technique could be employed to study thrombotic risks in a variety of disease states.