Nanoscale imaging using differential expansion microscopy
Nanoscale imaging using differential expansion microscopy
复制标题
DOI：
10.1007/s00418-020-01869-7
复制
发表时间：
2020-03-19
影响因子：
2.3
通讯作者：
Jena, Bhanu P.
中科院分区：
文献类型：
--
作者：
Pernal, Sebastian P.;Liyanaarachchi, Asiri;Jena, Bhanu P.
活检组织的免疫电子显微镜检查的昂贵和耗时的方法继续作为诊断病理学的金标准。扩展显微镜（ExM）的新方法的最近发展，能够四倍的横向扩展的生物标本的形态学检查，在约70 nm的横向分辨率，使用普通的衍射限制光学显微镜，是在细胞成像的一个重大进展。在这里，我们报告（1）一个优化的固定方案，以保留细胞形态，同时获得最佳的扩张，（2）一个ExM程序高达8倍的横向和超过500倍的体积扩张，（3）证明ExM是各向异性或组织之间的差异，细胞器和细胞器本身内的域，以及（4）应用图像分析和机器学习（ML）方法来精确地评估差异扩展的细胞结构。我们将这种与ML相结合的增强ExM方法称为差示膨胀显微镜（DiExM），适用于对纳米尺度的生物样本进行分析。DiExM在从病理标本载玻片中精确、快速和廉价地诊断疾病方面具有很大的潜力。
Expensive and time-consuming approaches of immunoelectron microscopy of biopsy tissues continues to serve as the gold-standard for diagnostic pathology. The recent development of the new approach of expansion microscopy (ExM) capable of fourfold lateral expansion of biological specimens for their morphological examination at approximately 70 nm lateral resolution using ordinary diffraction limited optical microscopy, is a major advancement in cellular imaging. Here we report (1) an optimized fixation protocol for retention of cellular morphology while obtaining optimal expansion, (2) an ExM procedure for up to eightfold lateral and over 500-fold volumetric expansion, (3) demonstrate that ExM is anisotropic or differential between tissues, cellular organelles and domains within organelles themselves, and (4) apply image analysis and machine learning (ML) approaches to precisely assess differentially expanded cellular structures. We refer to this enhanced ExM approach combined with ML as differential expansion microscopy (DiExM), applicable to profiling biological specimens at the nanometer scale. DiExM holds great promise for the precise, rapid and inexpensive diagnosis of disease from pathological specimen slides.