Rapid three-dimensional analysis of renal biopsy sections by low vacuum scanning electron microscopy.
Rapid three-dimensional analysis of renal biopsy sections by low vacuum scanning electron microscopy.
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通过低真空扫描电子显微镜对肾活检切片进行快速三维分析。
DOI：
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发表时间：
2011
影响因子：
--
通讯作者：
T. Naguro
中科院分区：
文献类型：
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作者：
S. Inaga;Masako Kato;S. Hirashima;C. Munemura;Sinichi Okada;T. Kameie;T. Katsumoto;H. Nakane;Kéiichi Tanaka;K. Hayashi;T. Naguro
肾活检石蜡切片检查低真空扫描电子显微镜（LVSEM）在背散射电子（BSE）模式，一种新的方法，用于快速病理分析，允许详细和有效的三维观察肾小球。在本研究中，对已经通过光学显微镜（LM）诊断为伊加肾病、轻微肾小球异常和膜性肾小球肾炎（GN）的肾脏样本进行快速处理。将载玻片上活检样本的未染色石蜡切片脱蜡，用铂蓝（Pt蓝）或高碘酸银-六亚甲基四胺（PAM）染色，并用LVSEM直接观察。在50 - 18000倍的任意放大倍数下，立即对整个切片进行概述，并对单个肾小球进行详细观察。同时显示肾小球切面和表面切面。在铂蓝染色切片上，由于BSE信号的产生量不同，可以区分足细胞、内皮细胞、系膜和肾小球基底膜（GBM），并研究每个样本的病理特征。足细胞的足突和不同厚度的GBM的异常表面外观的三维显示，在光镜和透射电子显微镜下难以观察的功能。膜性GN中PAM阳性GBM的改变在LVSEM下通过未去除细胞的覆盖细胞在高倍放大下清楚地可视化。在GBM的侧视图中，不仅可以清楚地看到明显的棘突形成，还可以看到轻微的突起。GBM的正面视图显示火山口状或孔状结构。因此，LVSEM有望提供一种新的方法，人类肾小球疾病的病理诊断，使用传统的肾活检切片。
Renal biopsy paraffin sections were examined by low vacuum scanning electron microscopy (LVSEM) in the backscattered electron (BSE) mode, a novel method for rapid pathological analysis which allowed detailed and efficient three-dimensional observations of glomeruli. Renal samples that had been already diagnosed by light microscopy (LM) as exhibiting IgA nephropathy, minor glomerular abnormalities, and membranous glomerulonephritis (GN) were rapidly processed in the present study. Unstained paraffin sections of biopsy samples on glass slides were deparaffinized, stained with platinum blue (Pt-blue) or periodic acid silver-methenamine (PAM), and directly observed with a LVSEM. Overviews of whole sections and detailed observations of individual glomeruli were immediately performed at arbitrary magnifications between ×50 to ×18,000. Cut surface views and surface views of glomeruli were demonstrated at the same time. On Pt-blue-stained sections, podocytes, endothelia, mesangium, and glomerular basement membranes (GBMs) could be distinguished due to the different yields of BSE signals, and pathological features were investigated in every sample. The abnormal surface appearances of podocytes with foot processes and the varying thicknesses of GBM were revealed three-dimensionally, features difficult to observe under LM and transmission electron microscopy. PAM-positive GBM alterations in membranous GN were distinctly visualized through overlying cells without cell removal under LVSEM at high magnification. Not only prominent spike formation but also slight protrusions were clearly revealed in the side views of GBM. Crater-like or hole-like structures were shown in the en face views of GBM. Accordingly, LVSEM is expected to provide a novel approach to the pathological diagnosis of human glomerular diseases using conventional renal biopsy sections.
影响因子：
2
作者：
PrestonJr,K;Joe,B;Siderits,R;Welling,J
通讯作者：
Welling,J