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用改良的切片冷冻干燥放射自显影技术测定氚标记对氨基马尿酸的肾内分布。

The intrarenal distribution of tritiated para-aminohippuric acid determined by a modified technique of section freeze-dry radioautography.

作者信息

Bordier B, Ornstein L, Weeden R P

出版信息

J Cell Biol. 1970 Sep;46(3):518-32. doi: 10.1083/jcb.46.3.518.

Abstract

Section freeze-dry radioautography has been used to examine the intrarenal distribution of a water-soluble organic acid (para-aminohippuric acid (PAH-3H)) under constant-infusion, steady-state conditions in mouse and rat kidney in vivo. The technique described here has the following advantages: (a) Sectioning and freeze-drying are accomplished in a closed cryostat at temperatures below -40 degrees C; (b) Handling of the section is facilitated by mounting of the section-to-be on adhesive-coated Saran Wrap prior to cutting; (c) Unembedded freeze-dried sections are attached to photographic film at ambient temperature in the dark room; (d) Fixation follows completion of radioautographic exposure and precedes photographic development; (e) Permanent close contact is maintained between tissue and film. Morphologic preservation compared favorably with that obtained by optimal fixation techniques, which, however, permit diffusion. Cellular accumulation of PAH-3H during secretion was demonstrated in the proximal tubule under steady-state conditions in vivo. The cellular concentration of PAH-3H was uniform throughout the length of the proximal tubule in mouse and rat kidney.

摘要

在体内小鼠和大鼠肾脏处于持续输注、稳态条件下,采用切片冷冻干燥放射自显影术来检测一种水溶性有机酸(对氨基马尿酸(PAH-3H))的肾内分布。此处描述的技术具有以下优点:(a)切片和冷冻干燥在温度低于-40℃的封闭低温恒温器中完成;(b)在切割前,通过将待切片安装在涂有粘合剂的莎纶薄膜上便于对切片进行处理;(c)未包埋的冷冻干燥切片在暗室中于室温下附着于摄影胶片;(d)固定在放射自显影曝光完成后、摄影显影之前进行;(e)组织与胶片之间保持永久紧密接触。形态学保存与通过最佳固定技术获得的结果相比具有优势,然而,最佳固定技术会允许扩散。在体内稳态条件下,近端小管中显示出PAH-3H在分泌过程中的细胞内积聚。在小鼠和大鼠肾脏近端小管全长中,PAH-3H的细胞浓度是均匀的。

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