title: Visualization of leukocyte transendothelial and interstitial migration using reflected light oblique transillumination in intravital video microscopy.
abstract: Dynamic visualization of the intravascular events leading to the extravasation of leukocytes into tissues by intravital microscopy has significantly expanded our understanding of the underlying molecular processes. In contrast, the detailed observation of leukocyte transendothelial and interstitial migration in vivo has been hampered by the poor image contrast of cells within turbid media that is obtainable by conventional brightfield microscopy. Here we present a microscopic method, termed reflected light oblique transillumination microscopy, that makes use of the optical interference phenomena generated by oblique transillumination to visualize subtle gradients of refractive indices within tissues for enhanced image contrast. Using the mouse cremaster muscle, we demonstrate that this technique makes possible the reliable quantification of extravasated leukocytes as well as the characterization of morphological phenomena of leukocyte transendothelial and interstitial migration.
authors.affiliation: Institute for Surgical Research, University of Munich, Munich, Germany.
authors.forename: Thorsten R
authors.initials: TR
authors.lastname: Mempel
authors.forename: Christian
authors.initials: C
authors.lastname: Moser
authors.forename: Joerg
authors.initials: J
authors.lastname: Hutter
authors.forename: Wolfgang M
authors.initials: WM
authors.lastname: Kuebler
authors.forename: Fritz
authors.initials: F
authors.lastname: Krombach
mesh_terms: D000818:Animals; D002450:Cell Communication; D002465:Cell Movement; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D007962:Leukocytes / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018715:Microscopy, Video; D014172:Transillumination