title: Calcium-imaging with Fura-2 in isolated cerebral microvessels.
abstract: Cerebral microvessels were isolated from rat hippocampus with a modified, mild collagenase digestion and loaded with the calcium fluoroprobe, Fura-2. Using a fluorescence microscope with quartz optics and an image analyser, it was possible to measure the intracellular concentration of free calcium ions [Ca2+]i in single microvessels for the first time. A resting level of (90 +/- 28) nmol/l (+/- SD, n = 24) was calculated which immediately rose after ionomycin application. The temporal resolution for [Ca2+]i of our set-up was 0.7 s. By image processing, sequences of digitized fluorescence images of single microvessels were colour-coded in terms of [Ca2+]i with a spatial resolution up to 1.5 microns (pixel size). Both the temporal and spatial resolution make our system suitable for investigation of calcium-mobilizing receptors of the blood-brain barrier.
authors.affiliation: Institute of Neuropathology, University of Copenhagen, Denmark.
authors.forename: J
authors.initials: J
authors.lastname: Hess
authors.forename: C V
authors.initials: CV
authors.lastname: Jensen
authors.forename: N H
authors.initials: NH
authors.lastname: Diemer
mesh_terms: D000818:Animals; D001572:Benzofurans; D002118:Calcium / Q000378:metabolism*; D005456:Fluorescent Dyes; D016257:Fura-2; D006624:Hippocampus / Q000098:blood supply*; D008833:Microcirculation / Q000166:cytology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D051381:Rats
chemical_list: D001572:Benzofurans; D005456:Fluorescent Dyes; D002118:Calcium; D016257:Fura-2