Rapid fluorescence-based measurement of neutrophil migration in vitro.
We have standardized a new chemotaxis chamber that uses fluorescence as the cellular marker for the measurement of leukocyte migration in vitro in disposable 96-well microplates. This new fluorescence-based assay is a robust assay because filter pore size, cell density, filter composition, and filter thickness do not affect PMN migration towards interleukin-8 or the complement fragment, C5a. When compared to two separate chemotaxis assays in which the migrated cells are counted visually, the fluorescence-based assay was more rapid, less labor intensive, and more sensitive. This new assay is a significant advance in the measurement of leukocyte migration in vitro.
Frevert
C W
CW
Medical Research Service of the Seattle VA Medical Center, Department of Medicine, University of Washington School of Medicine, 98108, USA. cfrevert@u.washington.edu
Wong
V A
VA
Goodman
R B
RB
Goodwin
R
R
Martin
T R
TR
D002452:Cell Count; D002634:Chemotaxis, Leukocyte; D005069:Evaluation Studies as Topic; D005452:Fluoresceins; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D008851:Micropore Filters; D009504:Neutrophils / Q000502:physiology*; D011076:Polycarboxylate Cement; D011091:Polyesters; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors
D005452:Fluoresceins; D005456:Fluorescent Dyes; D011076:Polycarboxylate Cement; D011091:Polyesters; C005506:polycarbonate; C007740:fluorexon