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Review of expansion microscopy combined with advanced imaging modalities

2025/11/18 by Natalie Woo, Claire M. Brown · 1 voice
Biochemistry, Genetics and Molecular Biology · Engineering · Physics and Astronomy · #Advanced Fluorescence Microscopy Techniques #Force Microscopy Techniques and Applications #Near-Field Optical Microscopy

paper · pdf · doi:10.1111/jmi.70048

openalex created_date 2025/11/18 · openalex publication_date 2025/11/18 · openalex updated_date 2026/08/01

Abstract

Expansion microscopy (ExM) is a powerful high-resolution imaging technique that enhances the spatial resolution of conventional light microscopy by physically enlarging biological specimens by embedding and cross-linking them in a swellable polymer network. This review explores the combination of ExM with commonly used advanced fluorescence imaging modalities, including light sheet fluorescence microscopy (LSFM), stimulated emission depletion (STED), structured illumination microscopy (SIM), single-molecule localisation microscopy (SMLM), and computational super-resolution radial fluctuations (SRRF) to push the boundaries of achievable resolution in biological imaging. By integrating ExM with these optical and analytical approaches, researchers can visualise subcellular structures and molecular complexes with unprecedented clarity, enabling the study of intricate biological processes that are otherwise inaccessible with conventional light microscopy methods. The review covers the theoretical resolutions attainable with each combined technique, example biological questions they can address, and key considerations for optimising their use. Together, these advancements offer novel insights into nanoscale cellular and subcellular structures, opening new avenues for exploration in fields such as neuroscience, cancer research, and developmental biology.

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