PMID- 32681506
OWN - NLM
STAT- MEDLINE
DCOM- 20210312
LR  - 20210312
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2161
DP  - 2020
TI  - Simultaneous Detection of RNAs and Proteins with Subcellular Resolution.
PG  - 59-73
LID - 10.1007/978-1-0716-0680-3_6 [doi]
AB  - We describe the detailed methods of "immunoFISH" to analyze the expression level 
      and the spatial localization of RNA transcripts and proteins on cultured cells 
      and formal-fixed, paraffin-embedded (FFPE) tissue sections. On cultured cells, we 
      detect specific transcripts using the Stellaris fluorescence in situ 
      hybridization (FISH) probes labeled with fluorophores that target multiple 
      regions along the desired transcripts and proteins combining the 
      immunofluorescent staining. On FFPE tissue sections, we use the RNAscope FISH 
      probes, modified branched DNA (bDNA) probes to amplify the RNA signals, followed 
      by immunofluorescent staining for protein detection. The abundance, composition, 
      and spatial distribution are determined by signals from fluorescently labeled 
      proteins and individual transcripts of images acquired using high-resolution 
      fluorescence microscopy.
FAU - Kwon, Sunjong
AU  - Kwon S
AD  - OHSU Center for Spatial Systems Biomedicine, Oregon Health and Sciences 
      University, Portland, OR, USA. kwons@ohsu.edu.
AD  - Department of Biomedical Engineering, Oregon Health and Sciences University, 
      Portland, OR, USA. kwons@ohsu.edu.
FAU - Chin, Koei
AU  - Chin K
AD  - OHSU Center for Spatial Systems Biomedicine, Oregon Health and Sciences 
      University, Portland, OR, USA.
AD  - Department of Biomedical Engineering, Oregon Health and Sciences University, 
      Portland, OR, USA.
FAU - Nederlof, Michel
AU  - Nederlof M
AD  - Quantitative Imaging Systems, Inc., Pittsburgh, PA, USA.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Proteins)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Animals
MH  - Cell Line, Tumor
MH  - Cells, Cultured
MH  - Fluorescent Dyes/chemistry
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods/standards
MH  - Limit of Detection
MH  - Proteins/metabolism
MH  - RNA, Messenger/metabolism
MH  - Staining and Labeling/methods
MH  - Tissue Embedding/methods
MH  - Tissue Fixation/methods
OTO - NOTNLM
OT  - Immunofluorescent
OT  - Subcellular localization
OT  - immunoFISH
OT  - smFISH
EDAT- 2020/07/19 06:00
MHDA- 2021/03/13 06:00
CRDT- 2020/07/19 06:00
PHST- 2020/07/19 06:00 [entrez]
PHST- 2020/07/19 06:00 [pubmed]
PHST- 2021/03/13 06:00 [medline]
AID - 10.1007/978-1-0716-0680-3_6 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2020;2161:59-73. doi: 10.1007/978-1-0716-0680-3_6.