Ravnic Dino J, Jiang Xiaoqun, Wolloscheck Tanja, Pratt Juan P, Huss Harold, Mentzer Steven J, Konerding Moritz A
Laboratory of Immunophysiology, Department of Surgery, Brigham and Women's Hospital, Harvard Surgical Research Laboratories, Harvard Medical School, Room 259, 75 Francis Street, Boston, MA 02115, USA.
Microvasc Res. 2005 Jul;70(1-2):90-6. doi: 10.1016/j.mvr.2005.06.002. Epub 2005 Aug 10.
Flexible approaches to defining microvessel morphometry are useful in the study of both acute and chronic structural changes of the microcirculation. In this report, we examined the utility of the intravascular infusion of lipophilic carbocyanine tracers in the structural assessment of the retina, skin, lung, and colon microcirculation. The microvessel labeling technique, here termed fluorescent vessel painting, involved the intravascular injection of sulfonated lipophilic carbocyanine tracers. The utility of vessel painting in morphometry was assessed using morphometric comparisons with corrosion casting and 2-dimensional and 3-dimensional scanning electron microscopy. The comparisons demonstrated that fluorescent vessel painting modestly overestimated the interbranch angles, interbranch distances, and vessel diameters of the 2D mucosal plexus of the colon. These differences were narrowed with the application of confocal microscopy. The advantages of fluorescence vessel painting included (1) the filling of all tissues including the relatively high resistance microvessels of the mouse skin, (2) the ability to use tissue counterstains such as DAPI, and (3) the prolonged stability of the lipophilic tracer after aldehyde fixation. These studies suggest the utility of fluorescent vessel painting as a complementary technique to corrosion casting in the morphometric study of the microcirculation.
灵活的微血管形态测量方法在研究微循环的急性和慢性结构变化中都很有用。在本报告中，我们研究了亲脂性碳菁染料示踪剂血管内灌注在视网膜、皮肤、肺和结肠微循环结构评估中的效用。微血管标记技术，这里称为荧光血管造影，涉及亲脂性磺化碳菁染料示踪剂的血管内注射。使用与铸型腐蚀以及二维和三维扫描电子显微镜的形态测量比较来评估血管造影在形态测量中的效用。比较结果表明，荧光血管造影略微高估了结肠二维黏膜丛的分支间角度、分支间距离和血管直径。应用共聚焦显微镜后，这些差异缩小了。荧光血管造影的优点包括：（1）能充盈所有组织，包括小鼠皮肤中相对高阻力的微血管；（2）能够使用如DAPI等组织复染剂；（3）醛固定后亲脂性示踪剂的稳定性延长。这些研究表明，荧光血管造影作为铸型腐蚀在微循环形态测量研究中的补充技术具有实用性。
