In vivo staining of neocortical astrocytes via the cerebral microcirculation using sulforhodamine B.
Staining and imaging glial cells in vivo while observing the microvasculature could help understand brain physiology, namely neuronal-glial-vascular communication. Two-photon excitation microscopy provides a means to monitor these interactions at the cellular level in living animals, but the cells of interest must be fluorescent. Injecting dyes intravenously is a rapid and quasi noninvasive method to stain cells in the brain. It necessitates that the dye is soluble in the blood plasma and crosses the blood brain barrier (BBB). We demonstrate here, using two-photon imaging, that sulforhodamine B (SRB) crosses the BBB and stains in vivo, specifically mouse astrocytes. This is confirmed by experiments on primary neurons and astrocytes cultures showing the preferential SRB staining of the latter. SRB is rapidly eliminated from the blood, which allows repeated injections in longitudinal studies.
Vérant
Pascale
P
Centre National de la Recherche Scientifique, Unite Mixte de Recherche 5588, Laboratoire de Spectrometrie Physique, Grenoble, F-38402 France.
Ricard
Clément
C
Serduc
Raphaël
R
Vial
Jean-Claude
JC
van der Sanden
Boudewijn
B
D000818:Animals; D001253:Astrocytes / Q000166:cytology*; D003287:Contrast Media / Q000008:administration & dosage; D007089:Image Enhancement / Q000379:methods*; D007275:Injections, Intravenous; D051379:Mice; D008833:Microcirculation; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D055806:Microvessels / Q000166:cytology*; D019579:Neocortex / Q000098:blood supply* / Q000166:cytology*; D012235:Rhodamines / Q000008:administration & dosage*; D013194:Staining and Labeling / Q000379:methods
D003287:Contrast Media; D012235:Rhodamines; C022027:lissamine rhodamine B