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JoVE Encyclopedia of Experiments
Cancer Research
高碘酸希夫或 PAS 染色:一种评估肾小球结构畸形的方法
高碘酸希夫或 PAS 染色:一种评估肾小球结构畸形的方法
Encyclopedia of Experiments
Cancer Research
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Encyclopedia of Experiments Cancer Research
Periodic Acid Schiff or PAS Staining: A Method to Assess the Structural Deformities of Glomerulus

高碘酸希夫或 PAS 染色:一种评估肾小球结构畸形的方法

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03:18 min
July 8, 2025
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Please note that some of the translations on this page are AI generated. Click here for the English version.

Transcript

肾小球是肾脏内复杂的毛细血管网络,用于过滤废物并去除血液中多余的液体。肾脏滤过率取决于这种特殊结构,可以使用过碘酸希夫或 PAS 染色剂进行检查。

首先,在载玻片上取石蜡化的肾皮层切片。用二甲苯处理载玻片。二甲苯分子溶解组织切片中的石蜡。接下来,将标本暴露在浓度降低的酒精中。酒精去除微量二甲苯,同时使组织切片重新水化,以便随后结合染色剂。

现在,将过碘酸添加到样品中。过碘酸将组织多糖中存在的二醇(例如糖原或粘蛋白)转化为醛。这些醛与希夫试剂(品红-亚硫酸组合)反应,形成洋红色染料,对组织切片进行染色。冲洗载玻片以去除多余的污渍。

冲洗后,加入铁苏木精,这是一种与 DNA 结合并将其染成紫色的核复染剂。清洗以去除多余的污渍。最后,用增加浓度的酒精对样品进行脱水,然后使用二甲苯,二甲苯用作澄清剂。此步骤有助于提高显微镜检查组织切片的对比度。

肾小球疾病可表现为基底膜增厚、毛细血管改变和与肾功能受损相关的异常细胞。

为了详细观察肾小球细胞和系膜基质,将固定石蜡包埋的肾皮质样品在37摄氏度下干燥1小时,然后用连续的二甲苯和下降乙醇浸泡脱蜡。将样品在蒸馏水中再水化 5 分钟,然后将载玻片在通风柜中的过碘酸溶液中孵育。

5 分钟后,在 100 毫升蒸馏水中用 3 次 5 分钟洗涤冲洗载玻片,然后在 Schiff 试剂中孵育 15 分钟。孵育结束时,在流动的自来水中清洗载玻片5分钟,并用苏木精复染3秒,然后在流动的自来水中彻底冲洗15分钟。

然后,用上升乙醇浸泡和两次连续 3 分钟的二甲苯浸泡使载玻片脱水。空气干燥后,载玻片可以用二甲苯基封片剂安装,以便在光学显微镜上以 400 倍放大倍率评估其肾小球结构。

Key Terms and Definitions

  • Periodic Acid Schiff Stain - Method used to visualize glomerular cells and the mesangial matrix.
  • PAS Stain Glomerulus - Specific application of PAS stain in kidney cortex study.
  • Acid Schiff Staining - Another name for the PAS staining process.
  • Periodic Acid Structure - Part of PAS stain, involved in revealing cellular structure in staining.
  • Glomerular Tuft - Structure in kidney cortex, analyzed using PAS stain.

Scientific Background

  • Periodic Acid Schiff Stain - Define and contextualize (e.g., periodic acid schiff stain).
  • Key Concepts - Summarize principles (e.g., staining theory).
  • Underlying Mechanisms - Short description of PAS staining process.
  • Connect to Experiment - Brief rationale linking periodic acid and schiff reagent to kidney cortex sample study.

Questions that this video will help you answer

  • What is the Periodic Acid Schiff Stain method and how it contributes to kidney cortex research?
  • How does the PAS staining process work?
  • What is the role of the Glomerular Tuft in PAS staining?

Applications and Relevance

  • Describe Practical Applications - PAS stain used for analyzing kidney structures (e.g., medical diagnosis).
  • Industry Impact - Benefits healthcare, particularly nephrology.
  • Societal Importance - Enables better understanding of kidney diseases (e.g., education).
  • Link to Scientific Advancements - Short description of innovations in staining techniques.

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