Optimization of Fluorescent Proteins and Techniques for In-resin Correlative Light and Electron Microscopy

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Microscopy and Microanalysis. 2020 Jul;26:1036–1039. doi: 10.1017/S143192762001675X

Optimization of Fluorescent Proteins and Techniques for In-resin Correlative Light and Electron Microscopy Looger Lab

Paez-Segala M, Wang Y, Iyer N, Li W, Rivlin P, Looger L

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Abstract

Correlative light and electron microscopy (CLEM) combines the power of electron microscopy, with its excellent resolution and contrast, with that of fluorescence imaging, which allows the staining of specific molecules, organelles, and cell populations. Fluorescence imaging is also readily compatible with live cells and behaving animals, facilitating real-time visualization of cellular processes, potentially followed by electron microscopic reconstruction. Super-resolution single-molecule localization microscopy is a relatively new modality that harnesses the ability of some fluorophores to photoconvert, through which localization precision better than Abbe’s diffraction limit is achieved through iterative high-resolution localization of single-molecule emitters. Here we describe our lab’s recent progress in the development of reagents and techniques for super-resolution single-molecule localization CLEM and their applications to biological problems.

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