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Methods. 2016 Apr 1;98:158-165. doi: 10.1016/j.ymeth.2015.11.007. Epub 2015 Nov 10.

Simultaneous multicolor detection of RNA and proteins using super-resolution microscopy.

Author information

1
RNA Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako 351-0198, Japan.
2
Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
3
RNA Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako 351-0198, Japan. Electronic address: nakagawas@riken.jp.

Abstract

A number of non-membranous cellular bodies have been identified in higher eukaryotes, and these bodies contain a specific set of proteins and RNAs that are used to fulfill their functions. The size of these RNA-containing cellular bodies is usually on a submicron scale, making it difficult to observe fine structures using optical microscopy due to the diffraction limitation of visible light. Recently, microscope companies have released super-resolution microscopes that were developed using different principles, enabling the observation of sub-micron structures not resolvable in conventional fluorescent microscopy. Here, we describe multi-color fluorescent in situ hybridization techniques optimized for the simultaneous detection of RNA and proteins using super-resolution microscopy, namely structured illumination microscopy (SIM).

KEYWORDS:

Cy2; In situ hybridization; Structured illumination microscopy (SIM); Super-resolution microscopy; TDE

PMID:
26564236
DOI:
10.1016/j.ymeth.2015.11.007
[Indexed for MEDLINE]
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