Feature-rich covalent stains for super-resolution and cleared tissue fluorescence microscopy.

Science Advances
Chenyi MaoJoshua C Vaughan

Abstract

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.

References

Jan 1, 1975·Histochemistry·F M WeinblattA M Seligman
Jun 1, 1990·Molecular and Biochemical Parasitology·D E HillJ F Urban
Mar 1, 1993·Bioconjugate Chemistry·R B MujumdarA S Waggoner
May 22, 2010·Proceedings of the National Academy of Sciences of the United States of America·Jeremy M BaskinCarolyn R Bertozzi
Jun 17, 2011·Journal of Proteome Research·Linn FagerbergEmma Lundberg
Feb 1, 2012·Nature Methods·Ulrike SchnellBen N G Giepmans
Dec 14, 2012·Proceedings of the National Academy of Sciences of the United States of America·Brian J BeliveauChao-ting Wu
Jan 17, 2015·Science·Fei ChenEdward S Boyden
Feb 6, 2015·Nature·Andrew Bradbury, Andreas Plückthun
Jul 18, 2015·Cell·Douglas S Richardson, Jeff W Lichtman
Apr 12, 2016·Nature Methods·Tyler J ChozinskiJoshua C Vaughan
Jun 10, 2016·ELife·Daniel N ItzhakGeorg Hh Borner
Jun 15, 2016·Annual Review of Cell and Developmental Biology·Kazuki TainakaHiroki R Ueda
Aug 5, 2016·Journal of the American Society of Nephrology : JASN·Anika KlingbergMatthias Gunzer
Sep 7, 2017·Nature Reviews. Molecular Cell Biology·Steffen J SahlStefan Jakobs
Dec 16, 2017·Kidney International·David Unnersjö-JessHjalmar Brismar
Jan 4, 2019·Nature Cell Biology·Lothar SchermellehGregor P C Drummen
Feb 10, 2019·Journal of Biomedical Optics·Weisi XieJonathan T C Liu
Feb 26, 2019·Database : the Journal of Biological Databases and Curation·Swagata PahariEmil Alexov
Mar 21, 2019·Biomedical Optics Express·Ye ChenJonathan T C Liu
Jul 6, 2019·Nature Communications·Adam K GlaserJonathan T C Liu
May 2, 2020·ELife·Chih-Chieh Jay YuEdward S Boyden

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Citations

Aug 2, 2020·Nature Communications·Ons M'Saad, Joerg Bewersdorf
Jan 19, 2021·Anatomical Science International·Yutaro Kashiwagi, Shigeo Okabe
Feb 17, 2021·Nature Biomedical Engineering·Jonathan T C LiuAnant Madabhushi

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Datasets Mentioned

BETA
GM130

Methods Mentioned

BETA
fluorescence microscopy
super-resolution microscopy
light microscopy
confocal microscopy
dissection

Key Resources (RRID) Mentioned

Addgene_54108
Addgene_15108

Software Mentioned

FLARE

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