TANDEM SCANNING CONFOCAL MICROSCOPY (TSCM) OF NORMAL AND ISCHEMIC LIVING KIDNEYS
TANDEM SCANNING CONFOCAL MICROSCOPY (TSCM) OF NORMAL AND ISCHEMIC LIVING KIDNEYS
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DOI：
10.1002/aja.1001910110
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发表时间：
1991-05-01
影响因子：
--
通讯作者：
JESTER, JV
中科院分区：
文献类型：
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作者：
ANDREWS, PM;PETROLL, WM;JESTER, JV
串联扫描共聚焦显微镜（TSCM）允许以非侵入性方式光学切片并记录活体器官和组织的实时图像。在本文中，我们将提出一些初步的TSCM观察包膜下肾单位在生活中，完整的肾脏Munich-Wistar大鼠和评估肾单位的反应暂时缺血和静脉输注甘露醇。用Inactin麻醉大鼠，并进行剖腹手术以暴露肾脏。使用配备20倍水浸物镜的TSCM，我们通过完整的肾包膜进行光学切片，并记录活的包膜下肾小球和肾小管的实时图像。近端小管刷状缘具有高度反射性，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穿过管周毛细血管和单个肾小球毛细血管环路。荧光过滤器的地方，静脉注射羧基荧光素被认为是通过肾小球毛细血管环路，然后逐步通过不同的段的肾小管。肾动脉结扎导致近端小管细胞迅速肿胀进入管腔，微绒毛刷状缘反射性丧失，以及小管周围毛细血管间隙闭合。在释放结扎线时，近端小管腔再次变得开放，通常突然打开，并以拉链样的方式沿着小管的长度。静脉输注甘露醇增加肾小球滤过率导致肾小管管腔和周长尺寸增加。甘露醇也作为一种有效的不渗透渗透剂，并防止大多数细胞肿胀，否则会出现肾缺血。
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.