Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/70868
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Type: Journal article
Title: Combined Boyden-flow cytometry assay improves quantification and provides phenotypification of leukocyte chemotaxis
Author: Gomez-Lopez, Nardhy Yadira
Vadillo-Ortega, Felipe
Estrada-Gutierrez, Guadalupe
Citation: PLoS One, 2011; 6(12):e28771
Publisher: Public Library of Science
Issue Date: 2011
ISSN: 1932-6203
School/Discipline: Obstetrics and Gynaecology
Statement of
Responsibility: 
Nardhy Gomez-Lopez, Felipe Vadillo-Ortega and Guadalupe Estrada-Gutierrez
Abstract: Chemotaxis has been studied by classical methods that measure chemotactic and random motility responses in vitro, but these methods do not evaluate the total number and phenotype of migrating leukocytes simultaneously. Our objective was to develop and validate a novel assay, combined Boyden-flow cytometry chemotaxis assay (CBFCA), for simultaneous quantification and phenotypification of migrating leukocytes. CBFCA exhibited several important advantages in comparison to the classic Boyden chemotaxis assay (CBCA): 1) improved precision (intra-assay coefficients of variation (CVs): CBFCA-4.7 and 4.8% vs. CBCA-30.1 and 17.3%; inter-observer CVs: CBFCA-3.6% vs. CBCA 30.1%); 2) increased recovery of cells, which increased assay to provide increased sensitivity; 3) high specificity for determining the phenotype of migrating/attracted leukocytes; and 4) reduced performance time (CBFCA 120 min vs. CBCA 265 min). Other advantages of CBFCA are: 5) robustness, 6) linearity, 7) eliminated requirement for albumin and, importantly, 8) enabled recovery of migrating leukocytes for subsequent studies. This latter feature is of great benefit in the study of migrating leukocyte subsets. We conclude that the CBFCA is a novel and improved technique for experiments focused on understanding leukocyte trafficking during the inflammatory response.
Description: Extent: 8p.
Rights: Copyright: © 2011 Gomez-Lopez, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
RMID: 0020115324
DOI: 10.1371/journal.pone.0028771
Appears in Collections:Obstetrics and Gynaecology publications

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