PMID: 11909751Mar 23, 2002Paper

Enumeration of micronucleated CD71-positive human reticulocytes with a single-laser flow cytometer

Mutation Research
Stephen D DertingerC R Tometsko

Abstract

The extreme rarity of micronucleated reticulocytes (RETs) in the peripheral blood of non-splenectomized humans has precluded facile enumeration of these cells, as well as evaluation of this endpoint as an index of cytogenetic damage. In this report, we describe a high-throughput, single-laser flow cytometric system for scoring the incidence of micronuclei (MN) in newly formed human RETs. The procedure is based on an immunochemical reagent that differentially labels the most immature fraction of RETs from mature erythrocytes based on the expression level of the transferrin receptor (also known as CD71). The resolution of four erythrocyte populations (young RETs and mature erythrocytes, with and without MN) was achieved for human blood cells treated with phycoerythrin-conjugated anti-CD71, RNase, and either SYTOX Green or SYBR Green I nucleic acid dyes. Anti-glycophorin A labeling of erythroid cells (CyChrome conjugate) was also incorporated into the staining procedure to ensure that debris or other potential artifacts did not adversely impact the analyses. Instrument calibration procedures utilizing malaria-infected rodent erythrocytes were also developed, and are described. Using this analytical system, blood samples from 10 he...Continue Reading

References

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Jul 27, 2001·Environmental and Molecular Mutagenesis·D K TorousC R Tometsko

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Citations

Dec 24, 2003·Teratogenesis, Carcinogenesis, and Mutagenesis·Elizabeth D WagnerMichael J Plewa
Dec 4, 2003·Mutation Research·Stephen D DertingerCarol R Tometsko
Jun 1, 2006·Blood·Patricia A OnealJeffery L Miller
May 30, 2003·Toxicological Sciences : an Official Journal of the Society of Toxicology·Dorothea K TorousStephen D Dertinger
Oct 29, 2010·Mutagenesis·John A HeddleJames T MacGregor
Apr 23, 2013·Journal of Tissue Engineering and Regenerative Medicine·Colleen ByrnesJeffery L Miller
Nov 2, 2004·Environmental and Molecular Mutagenesis·Stephen D DertingerGeorge Bolcsfoldi
Sep 22, 2010·Mutation Research·Véronique ThybaudJan Van Benthem
Feb 15, 2005·Mutation Research·Vasily N DobrovolskyRobert H Heflich
Jan 1, 2015·Nature Protocols·Gabriel BalmusRebecca E McIntyre
Feb 23, 2012·Scientific Reports·Benoît MalleretLaurent Rénia
Oct 20, 2018·Biomédica : revista del Instituto Nacional de Salud·Lina Marcela BarreraMauricio Camargo
Mar 9, 2019·Tissue Engineering. Part C, Methods·Seo Hui KimEun Jung Baek
Jun 29, 2011·Journal of Toxicology and Environmental Health. Part a·Carla CostaJoão Paulo Teixeira
Dec 17, 2004·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Tal OfferBruce N Ames
Dec 20, 2020·Mutation Research. Reviews in Mutation Research·Richard J Albertini, Debra A Kaden

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