Plasma and Red Blood Cell in Focal Cerebral Ischemia: Differential Visualization with Double Fluorescence Technique
We recently developed a method to study the microvascular perfusion within the brain parenchyma using fluorescence-labeled plasma . The method was extended in the present study to visualize circulated plasma as well as red blood cells (RBC) simultaneously in the same brain slice . After centrifugation and repeated washing with glucose-saline-barbiturate buffer, RBC of normal gerbils were labeled with dichlorotriazinil aminofluorescein-2HCl (DTAF) at pH 9.0 for 1 h at room temperature. The gerbil plasma was similarly labeled with tetramethyl rhodamine isothiocyanate (TRITC). The RBC suspension and the plasma were mixed (4:1) to make up double-labeled blood. The posterior communicating artery of a gerbil was occluded [3), producing ischemia in the ipsilateral hippocampus and thalamus [4–6]. From 30 min to 3 h after occlusion, 0.5 ml of the double-labeled blood was injected in 5 s from the femoral vein. Thirty seconds after the injection, the brain was removed and fixed in 95% ethanol. Coronal sections, 25 μm thick, were mounted in glycerol and examined under the fluorescence microscope.
KeywordsCerebral Ischemia Focal Cerebral Ischemia Cereb Blood Flow Microvascular Perfusion Ipsilateral Hippocampus
Unable to display preview. Download preview PDF.
- 1.Yoshimine T, Kato A, Hayakawa T, Ushio Y, Mogami H (1987) Deterioration and recovery of microcirculation during cerebral ischemia and recirculation. J Cereb Blood Flow Metabol 7: S337Google Scholar
- 2.Yoshimine T, Hayakawa T, Mogami H (1988) Plasma and RBC circulation in focal cerebral ischemia. Differential visualization with double fluorescence technique. In: Tomita M, Sawada T, Naritomi H, Heiss W-D (eds) Excerpta Medica International Congress Series. Cerebral Hyperemia and Ischemia. From the standpoint of cerebral blood volume, Elsevier, Amsterdam, pp 65–72Google Scholar