Andrews P M, Petroll W M, Cavanagh H D, Jester J V
Department of Anatomy and Cell Biology, Georgetown University Medical Center, Washington, D.C. 20007.
Am J Anat. 1991 May;191(1):95-102. doi: 10.1002/aja.1001910110.
Tandem Scanning Confocal Microscopy (TSCM) allows one to section optically into and record real-time images of living organs and tissues in a noninvasive fashion. In this paper, we will present some initial TSCM observations of subcapsular nephrons in the living, intact kidneys of Munich-Wistar rats and evaluate the nephron's responses to temporary ischemia and to intravenous infusion of mannitol. The rats were anesthetized with Inactin and a laparotomy performed to expose the kidneys. Using a TSCM equipped with a 20 x water-immersion objective, we optically sectioned through the intact kidney capsule and recorded real-time images of living subcapsular glomeruli and uriniferous tubules. The proximal tubule brush border was highly reflective and allowed us to distinguish between the first and second segments of the proximal tubules as well as the distal tubules. Cellular elements of the blood could be seen passing rapidly through peritubular capillaries and individual glomerular capillary loops. With fluorescent filters in place, intravenously injected carboxyfluorescein was seen to pass through the glomerular capillary loops and then progressively through the different segments of the uriniferous tubules. Ligation of the renal artery resulted in rapid swelling of proximal tubule cells into the tubular lumens, loss of reflectiveness of the microvillous brush borders, and closure of the peritubular capillary spaces. Upon release of the ligature, the proximal tubule lumens again became patent, often opening up abruptly and in a zipper-like fashion down the length of the tubules. Increasing the glomerular filtration rate by intravenous infusion of mannitol resulted in increases in tubular luminal and perimeter dimensions. Mannitol also acted as an effective impermeant osmotic agent and prevented most of the cellular swelling which was otherwise seen in response to renal ischemia.
串联扫描共聚焦显微镜（TSCM）使人们能够以非侵入性方式对活体器官和组织进行光学切片并记录实时图像。在本文中，我们将展示对慕尼黑 - 威斯塔大鼠完整活体肾脏中被膜下肾单位的一些初步TSCM观察结果，并评估肾单位对暂时缺血和静脉输注甘露醇的反应。大鼠用安泰酮麻醉并进行剖腹手术以暴露肾脏。使用配备20倍水浸物镜的TSCM，我们对完整的肾被膜进行光学切片，并记录活体被膜下肾小球和肾小管的实时图像。近端小管刷状缘具有高反射性，使我们能够区分近端小管的第一段和第二段以及远端小管。可以看到血液中的细胞成分迅速通过肾小管周围毛细血管和单个肾小球毛细血管袢。在安装荧光滤光片后，可以看到静脉注射的羧基荧光素穿过肾小球毛细血管袢，然后逐渐穿过肾小管的不同节段。结扎肾动脉导致近端小管细胞迅速肿胀进入管腔，微绒毛刷状缘的反射性丧失，以及肾小管周围毛细血管间隙关闭。松开结扎线后，近端小管腔再次通畅，通常会突然以拉链状方式沿小管长度打开。通过静脉输注甘露醇增加肾小球滤过率导致管腔和周长尺寸增加。甘露醇还作为一种有效的非渗透性渗透剂，防止了大多数因肾缺血而出现的细胞肿胀。
