Tata D A, Anderson B J
Department of Psychology, State University of New York at Stony Brook, Stony Brook, New York, NY 11794-2500, USA.
J Neurosci Methods. 2002 Jan 30;113(2):199-206. doi: 10.1016/s0165-0270(01)00494-0.
Numerous physiological conditions as well as behavioral conditions have been shown to influence central nervous system vascular structure. Many of the methods used to investigate these structural alterations take advantage of the visibility of viscous substances (e.g. India ink in gelatin) perfused into the vasculature. The high viscosity of the solution, however, can cause incomplete vessel perfusion. The aim of the present study was to test whether or not capillaries seen in tissue perfused with fixative, embedded in celloidin and stained with Methylene Blue-Azure II (n=6) could be a useful alternative for the investigation of brain vascular structure. The method was compared to tissue from six rats perfused with India ink in gelatin and stained with cresyl violet. Qualitatively, vessels in the standard perfused tissue embedded in celloidin yielded clear vessels with stained pericytes. The two methods did not differ in branch point to cell ratio, length of individual capillaries, vessel length per mm(3), and capillary tortuosity. The capillary diameter was greater in the celloidin embedded tissue than in the India ink perfused tissue. Measuring the diameter between vessel walls appears to provide a more accurate measure than the widest distance between India ink pigments. Quantitative comparisons suggest that perfusion with standard fixative followed by embedding in celloidin provides vascular quantification comparable to that from India ink perfused tissue. The present method has several advantages, which include visualization of pericytes, increased probability of complete perfusion, clear view of cells that might otherwise be obscured by opaque vessels, and the possibility of using the alternate cerebral hemisphere for investigation of vascular ultrastructure.
众多生理状况以及行为状况已被证明会影响中枢神经系统的血管结构。许多用于研究这些结构改变的方法利用了灌注到脉管系统中的粘性物质（如明胶中的印度墨水）的可视性。然而，溶液的高粘度会导致血管灌注不完全。本研究的目的是测试在用固定剂灌注、火棉胶包埋并用亚甲蓝 - 天青II染色的组织中看到的毛细血管（n = 6）是否可以作为研究脑血管结构的有用替代方法。将该方法与六只用明胶中的印度墨水灌注并用甲酚紫染色的大鼠的组织进行比较。定性地说，火棉胶包埋的标准灌注组织中的血管产生了带有染色周细胞的清晰血管。两种方法在分支点与细胞比例、单个毛细血管长度、每立方毫米血管长度和毛细血管曲折度方面没有差异。火棉胶包埋组织中的毛细血管直径大于印度墨水灌注组织中的毛细血管直径。测量血管壁之间的直径似乎比测量印度墨水色素之间的最宽距离提供更准确的测量。定量比较表明，用标准固定剂灌注然后用火棉胶包埋提供的血管定量与印度墨水灌注组织的血管定量相当。本方法有几个优点，包括周细胞的可视化、完全灌注的可能性增加、对否则可能被不透明血管遮挡的细胞的清晰观察，以及使用对侧大脑半球研究血管超微结构的可能性。
