FLARE: Feature-Rich Covalent Stains for Super-Resolution and Cleared-Tissue Fluorescence Microscopy

Mouse small intestine stained via FLARE
Mao, C.; Lee, M. Y.; Jhan, J.-R.; Halpern, A. R.; Woodworth, M. A.; Glaser, A. K.; Chozinski, T. J.; Shin, L.; Pippin, J.W.; Shankland, S.J.; Liu, J. T. C.; Vaughan, J. C. Feature-Rich Covalent Stains for Super-Resolution and Cleared-Tissue Fluorescence Microscopy. Sci. Adv. 2020, 6, eaba4542.

Fluorescence microscopy is a workhorse tool in biomedical imaging but often poses substantial challenges to practitioners in achieving bright or uniform labeling. In addition, while antibodies are effective specific labels, their reproducibility is often inconsistent, and they are difficult to use when staining thick specimens. We report the use of conventional, commercially available fluorescent dyes for rapid and intense covalent labeling of proteins and carbohydrates in super-resolution (expansion) microscopy and cleared tissue microscopy. This approach, which we refer to as Fluorescent Labeling of Abundant Reactive Entities (FLARE), produces simple and robust stains that are modern equivalents of classic small-molecule histology stains. It efficiently reveals a wealth of key landmarks in cells and tissues under different fixation or sample processing conditions and is compatible with immunolabeling of proteins and in situ hybridization labeling of nucleic acids.

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