Plasma and Red Blood Cell in Focal Cerebral Ischemia: Differential Visualization with Double Fluorescence Technique

  • Toshiki Yoshimine
  • Toru Hayakawa
  • Heitaro Mogami


We recently developed a method to study the microvascular perfusion within the brain parenchyma using fluorescence-labeled plasma [1]. 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 [2]. 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.


Cerebral Ischemia Focal Cerebral Ischemia Cereb Blood Flow Microvascular Perfusion Ipsilateral Hippocampus 
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Copyright information

© Springer-Verlag Tokyo 1988

Authors and Affiliations

  • Toshiki Yoshimine
  • Toru Hayakawa
  • Heitaro Mogami
    • 1
  1. 1.Department of NeurosurgeryOsaka University Medical SchoolFukushima-ku, Osaka 553Japan

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