Modified labeling technique for in vivo visualization of platelets in the cerebral microcirculation of Mongolian gerbils

摘要: Activation of platelets induces interactions with platelets, endothelial cells, and leukocytes. In vivo observation these in the cerebral microcirculation is rare. The purpose present study was to develop a model enabling platelet kinetics microcirculation. Intravital fluorescence microscopy performed Mongolian gerbil. Platelets donor were labeled ex carboxyfluorescein diacetat-succinimidylester (CFDA-SE), providing long-term fluorescence. Platelet function tested by flow cytometric analysis analyzing platelet-endothelium interactions. Labeled stimulated adenosine diphosphate ADP (200 micromol/L) or thrombin (1000 U/L) showed aggregation cytrometric analysis, whereas unstimulated not aggregated. Irradiation brain surface after intravenous injection photosensitizing dye Photosan first induced rolling firm adherence on arteriolar venular endothelium, followed formation thrombus obstructing vessel. Quantitative (n x 100 microm(-1) min(-1)) before 6 mins irradiation 2.6+/-3.2 versus 29.0+/-28.9 rolling, 0.0+/-0.0 1.7+/-2.3 adherent arterioles, 3.9+/-3.3 36.6+/-20.9 13.6+/-8.9 venules. Thus, we conclude that labeling CFDA-SE does activate platelets. adhesion achieved platelet-specific stimulation such as ADP, irradiation. assessment physiologic pathophysiologic mechanisms can be this model.