Micro-Sieving: Isolation of Whole Glomeruli from a Single Renal Needle Biopsy Sample
Micro-Sieving: Isolation of Whole Glomeruli from a Single Renal Needle Biopsy Sample
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DOI：
10.1159/000320198
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发表时间：
2011-01-01
影响因子：
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通讯作者：
Kato, Tomohiro
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文献类型：
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作者：
Koitabashi, Kenichiro;Okamoto, Kazuki;Kato, Tomohiro
肾活检样本不仅对于肾小球肾炎的诊断很重要，而且对于其发病机制的研究也很重要。然而，对仅从针吸活检样本的肾小球中提取的蛋白质进行生化分析仍然很困难，因为样本含有肾小管和结缔组织等多种成分。即使是最近的显微解剖方法，在活检样本的切片中恢复肾小球，也没有完全解决这个困难，因为通过这种方法获得的蛋白质的量通常不足以进行蛋白质分析。为了克服这个问题，我们建立了一种简单但可靠的方法来从针吸活检样本中分离整个肾小球。通过这种称为“微筛分”的方法，我们能够在一小时内从单个针活检样本中平均分离出 50 多个肾小球。每个活检样本提取的肾小球蛋白的量平均为 23 μg。作为该方法的代表性应用，我们能够通过荧光二维电泳获得每位接受测试的肾小球肾炎患者的肾小球蛋白质谱。 “微筛分”可以广泛用作分析肾针活检样本中肾小球的基本技术。版权所有 (C) 2010 S. Karger AG，巴塞尔
Renal biopsy samples are important not only for the diagnosis of glomerulonephritis, but also for the investigation of its pathogenesis. However, it remains difficult to biochemically analyze proteins extracted solely from the glomeruli of needle biopsy samples, since the samples contain various components like renal tubules and connective tissue. Even a recent micro-dissection method, recovering the glomeruli in the sliced sections of the biopsy samples, has not fully solved the difficulty because the amount of obtainable proteins by this method is not usually enough for protein analysis. To overcome this problem, we established a simple but reliable method to isolate whole glomeruli from needle biopsy samples. By this method, termed 'micro-sieving', we were able to isolate on average more than 50 glomeruli from a single needle biopsy sample in an hour. The amount of the extracted glomerular proteins was on average 23 mu g per biopsy sample. As a representative use of this method, we were able to obtain a glomerular protein profile by fluorescent 2-dimensional electrophoresis for each of the tested patients with glomerulonephritis. 'Micro-sieving' can be used widely as a fundamental technique to analyze glomeruli in renal needle biopsy samples. Copyright (C) 2010 S. Karger AG, Basel