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Annual Review of Biomedical Engineering

Volume 22, 2020

Three-Dimensional Single-Molecule Localization Microscopy in Whole-Cell and Tissue Specimens

Abstract

Super-resolution microscopy techniques are versatile and powerful tools for visualizing organelle structures, interactions, and protein functions in biomedical research. However, whole-cell and tissue specimens challenge the achievable resolution and depth of nanoscopy methods. We focus on three-dimensional single-molecule localization microscopy and review some of the major roadblocks and developing solutions to resolving thick volumes of cells and tissues at the nanoscale in three dimensions. These challenges include background fluorescence, system- and sample-induced aberrations, and information carried by photons, as well as drift correction, volume reconstruction, and photobleaching mitigation. We also highlight examples of innovations that have demonstrated significant breakthroughs in addressing the abovementioned challenges together with their core concepts as well as their trade-offs.

Keyword(s): adaptive optics, Cramér–Rao lower bound, CRLB, fluorescence microscopy, light-sheet microscopy, super-resolution microscopy, tissue imaging
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/content/journals/10.1146/annurev-bioeng-060418-052203
2020-06-04
2024-04-19
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/content/journals/10.1146/annurev-bioeng-060418-052203
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Three-Dimensional Single-Molecule Localization Microscopy in Whole-Cell and Tissue Specimens
Annual Review of Biomedical Engineering 22, 155 (2020); https://doi.org/10.1146/annurev-bioeng-060418-052203
/content/journals/10.1146/annurev-bioeng-060418-052203
/content/journals/10.1146/annurev-bioeng-060418-052203
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