Correlated confocal and super-resolution imaging by VividSTORM

Barna László; Dudok Barna; Csapóné Miczán Vivien; Horváth András; László Zsófia Ilona; Katona István: Correlated confocal and super-resolution imaging by VividSTORM.
NATURE PROTOCOLS, 11 (1). pp. 163-183. ISSN 1754-2189 (2016)

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nprot.2016.002.pdf - Megjelent verzió
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Folyóiratcikk

Szerző azonosítók:

Név	ORCID	MTMT szerző azonosító
Barna László		10053111
Dudok Barna	0000-0001-5078-0090	230
Csapóné Miczán Vivien		10052396
Horváth András	0000-0001-5855-4186	10029872
László Zsófia Ilona		10055721
Katona István		10011502

Absztrakt (kivonat):

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.

Folyóirat címe:

NATURE PROTOCOLS

Megjelenés éve:

2016

Kötet:

Szám:

Oldalak:

pp. 163-183

ISSN:

1754-2189

Intézmény:

Pázmány Péter Katolikus Egyetem