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Nanoscale Imaging of Kidney Glomeruli Using Expansion Pathology

DOI:10.3389/fmed.2018.00322 期刊:Frontiers in Medicine 出版年份:2018 更新时间:2025-09-09 09:28:46
摘要: Kidney glomerular diseases, such as the minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS), and other nephrotic syndromes, are typically diagnosed or confirmed via electron microscopy. Although optical microscopy has been a vital tool to examine clinical specimens for diagnoses in pathology for decades, the optical resolution is constricted by the physical diffraction limit of the optical microscope, which prevents high-resolution investigation of subcellular anatomy, such as of the podocyte tertiary foot processes. Here, we describe a simple, fast, and inexpensive protocol for nanoscale optical imaging of kidney glomeruli. The protocol is based on Expansion Pathology (ExPath), a new principle of microscopy that overcomes optical diffraction limit by chemically embedding specimens into a swellable polymer and physically expanding it homogenously prior to imaging. Our method uses only commercially available reagents, a conventional fluorescence microscope and it can be applied to both fixed-frozen or formalin-fixed paraffin embedded (FFPE) tissue sections. It requires minimal operative experience in a wet lab, optical microscopy and imaging processing. Finally, we also discuss challenges, limitations and prospective applications for ExPath-based imaging of glomeruli.
作者: Octavian Bucur,Yongxin Zhao
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To describe a protocol for nanoscale optical imaging of kidney glomeruli using Expansion Pathology (ExPath), enabling the visualization of subcellular anatomy beyond the diffraction limit of conventional optical microscopy.

ExPath enables visualization of tertiary podocyte foot processes in kidney glomeruli using conventional optical microscopes, significantly lowering equipment requirements and potential costs for examining kidney biopsies with glomerular diseases. Future large-scale blinded studies using ExPath may streamline the diagnosis or confirmation of kidney glomerular diseases.

ExPath is not compatible with living specimens and is limited to fixed specimens. It allows investigation of up to 4-5 molecular targets with fluorescence microscopy. Imaging whole expanded tissue slides may take longer due to increased specimen volume and diluted fluorescence signals.

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