Correlative light and electron microscopy for human brain and other biological models.

IF 13.1 1区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Nature Protocols Pub Date : 2025-03-31 DOI:10.1038/s41596-025-01153-9

Notash Shafiei, Daniel Stӓhli, Domenic Burger, Marta Di Fabrizio, Lukas van den Heuvel, Jean Daraspe, Carolin Böing, Sarah H Shahmoradian, Wilma D J van de Berg, Christel Genoud, Henning Stahlberg, Amanda J Lewis

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引用次数: 0

Abstract

Correlative light and electron microscopy (CLEM) combines light microscopy, for identifying a target via genetic labels, dyes, antibodies and morphological features, with electron microscopy, for analyzing high-resolution subcellular ultrastructures. Here, we describe step-by-step instructions to perform a CLEM experiment, optimized for the investigation of ultrastructural features in human brain tissue. The procedure is carried out at room temperature and can be adapted to other human and animal tissue samples. The procedure requires 8 d to complete and includes the stages of sample fixation for optimal ultrastructural preservation, immunofluorescence staining, image acquisition and multimodal image correlation and is executable within standard electron microscopy laboratories. Serving as a critical tool for characterizing human tissue and disease models, room-temperature CLEM facilitates the identification and quantification of subcellular morphological features across brain regions.

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相关光学和电子显微镜用于人脑和其他生物模型。

相关光学和电子显微镜（CLEM）结合了光学显微镜，通过遗传标记，染料，抗体和形态特征识别目标，电子显微镜，用于分析高分辨率的亚细胞超微结构。在这里，我们描述一步一步的指令来执行CLEM实验，优化了人类脑组织的超微结构特征的调查。该程序在室温下进行，可以适用于其他人类和动物组织样本。该过程需要8天完成，包括样品固定以获得最佳超微结构保存、免疫荧光染色、图像采集和多模态图像相关等阶段，可在标准电子显微镜实验室中执行。作为表征人体组织和疾病模型的关键工具，室温CLEM有助于识别和量化跨脑区域的亚细胞形态学特征。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Nature Protocols 生物-生化研究方法

CiteScore

29.10

自引率

0.70%

发文量

128

审稿时长

4 months

期刊介绍： Nature Protocols focuses on publishing protocols used to address significant biological and biomedical science research questions, including methods grounded in physics and chemistry with practical applications to biological problems. The journal caters to a primary audience of research scientists and, as such, exclusively publishes protocols with research applications. Protocols primarily aimed at influencing patient management and treatment decisions are not featured. The specific techniques covered encompass a wide range, including but not limited to: Biochemistry, Cell biology, Cell culture, Chemical modification, Computational biology, Developmental biology, Epigenomics, Genetic analysis, Genetic modification, Genomics, Imaging, Immunology, Isolation, purification, and separation, Lipidomics, Metabolomics, Microbiology, Model organisms, Nanotechnology, Neuroscience, Nucleic-acid-based molecular biology, Pharmacology, Plant biology, Protein analysis, Proteomics, Spectroscopy, Structural biology, Synthetic chemistry, Tissue culture, Toxicology, and Virology.