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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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.