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Micro-sieving: isolation of whole glomeruli from a single renal needle biopsy sample.

机译:微筛:从单个肾穿刺活检样本中分离出整个肾小球。

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摘要

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 mug 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.
机译:肾活检标本不仅对肾小球肾炎的诊断很重要,而且对于其发病机理的研究也很重要。但是,由于生化样品中含有肾小管和结缔组织等各种成分,因此仅从针状活检样品的肾小球中提取的蛋白质进行生化分析仍然很困难。即使是最近的显微解剖方法,在活检样本的切片部分中回收肾小球,也不能完全解决这一难题,因为通过这种方法获得的蛋白质数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而可靠的方法来从穿刺活检样本中分离出整个肾小球。通过这种称为“微筛分”的方法,我们能够在一小时内从单针活检样本中平均分离出50多个肾小球。每个活检样本平均提取的肾小球蛋白量为23马克杯。作为该方法的代表用途,我们能够通过荧光二维电泳为每位接受测试的肾小球肾炎患者获得肾小球蛋白谱。 “微筛”可广泛用作分析肾针活检样本中肾小球的基本技术。

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