Correlated confocal and super-resolution imaging by VividSTORM

László Barna, Barna Dudok, Vivien Miczán, András Horváth, Zsófia I. László, I. Katona

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Single-molecule localization microscopy (SMLM) is rapidly gaining popularity in the life sciences as an efficient approach to visualize molecular distribution with nanoscale precision. However, it has been challenging to obtain and analyze such data within a cellular context in tissue preparations. Here we describe a 5-d tissue processing and immunostaining procedure that is optimized for SMLM, and we provide example applications to fixed mouse brain, heart and kidney tissues. We then describe how to perform correlated confocal and 3D-superresolution imaging on these sections, which allows the visualization of nanoscale protein localization within labeled subcellular compartments of identified target cells in a few minutes. Finally, we describe the use of VividSTORM (http://katonalab.hu/index.php/vividstorm), an open-source software for correlated confocal and SMLM image analysis, which facilitates the measurement of molecular abundance, clustering, internalization, surface density and intermolecular distances in a cell-specific and subcellular compartment-restricted manner. The protocol requires only basic skills in tissue staining and microscopy.

Original languageEnglish
Pages (from-to)163-183
Number of pages21
JournalNature Protocols
Volume11
Issue number1
DOIs
Publication statusPublished - Jan 3 2016

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Microscopic examination
Tissue
Imaging techniques
Molecules
Biological Science Disciplines
Image analysis
Cluster Analysis
Microscopy
Brain
Software
Visualization
Staining and Labeling
Kidney
Network protocols
Processing
Single Molecule Imaging
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Barna, L., Dudok, B., Miczán, V., Horváth, A., László, Z. I., & Katona, I. (2016). Correlated confocal and super-resolution imaging by VividSTORM. Nature Protocols, 11(1), 163-183. https://doi.org/10.1038/nprot.2016.002

Correlated confocal and super-resolution imaging by VividSTORM. / Barna, László; Dudok, Barna; Miczán, Vivien; Horváth, András; László, Zsófia I.; Katona, I.

In: Nature Protocols, Vol. 11, No. 1, 03.01.2016, p. 163-183.

Research output: Contribution to journalArticle

Barna, L, Dudok, B, Miczán, V, Horváth, A, László, ZI & Katona, I 2016, 'Correlated confocal and super-resolution imaging by VividSTORM', Nature Protocols, vol. 11, no. 1, pp. 163-183. https://doi.org/10.1038/nprot.2016.002
Barna L, Dudok B, Miczán V, Horváth A, László ZI, Katona I. Correlated confocal and super-resolution imaging by VividSTORM. Nature Protocols. 2016 Jan 3;11(1):163-183. https://doi.org/10.1038/nprot.2016.002
Barna, László ; Dudok, Barna ; Miczán, Vivien ; Horváth, András ; László, Zsófia I. ; Katona, I. / Correlated confocal and super-resolution imaging by VividSTORM. In: Nature Protocols. 2016 ; Vol. 11, No. 1. pp. 163-183.
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