PMID- 38502485
OWN - NLM
STAT- MEDLINE
DCOM- 20240321
LR  - 20240321
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2784
DP  - 2024
TI  - Multiplexed Immunofluorescence and Single-Molecule RNA Fluorescence In Situ 
      Hybridization in Mouse Skeletal Myofibers.
PG  - 163-176
LID - 10.1007/978-1-0716-3766-1_11 [doi]
AB  - RNA fluorescence in situ hybridization (FISH) is a powerful method to determine 
      the abundance and localization of mRNA molecules in cells. While modern RNA FISH 
      techniques allow quantification at single molecule resolution, most methods are 
      optimized for mammalian cell culture and are not easily applied to in vivo tissue 
      settings. Single-molecule RNA detection in skeletal muscle cells has been 
      particularly challenging due to the thickness and high autofluorescence of adult 
      muscle tissue and a lack of in vitro models for mature muscle cells (myofibers). 
      Here, we present a method for isolation of adult myofibers from mouse skeletal 
      muscle and detection of single mRNA molecules and proteins using multiplexed RNA 
      FISH and immunofluorescence.
CI  - (c) 2024. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Denes, Lance T
AU  - Denes LT
AD  - Institute for Systems Genetics, NYU Grossman School of Medicine, New York, NY, 
      USA. lance.denes@nyulangone.org.
FAU - Kelley, Chase P
AU  - Kelley CP
AD  - Department of Molecular Genetics and Microbiology, University of Florida, 
      Gainesville, FL, USA.
FAU - Wang, Eric T
AU  - Wang ET
AD  - Department of Molecular Genetics and Microbiology, University of Florida, 
      Gainesville, FL, USA.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 63231-63-0 (RNA)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Mice
MH  - Animals
MH  - *RNA/genetics/metabolism
MH  - In Situ Hybridization, Fluorescence/methods
MH  - *Muscle Fibers, Skeletal/metabolism
MH  - RNA, Messenger/genetics/metabolism
MH  - Fluorescent Antibody Technique
MH  - Muscle, Skeletal
MH  - Mammals
OTO - NOTNLM
OT  - Fluorescence imaging
OT  - Myofiber
OT  - RNA
OT  - RNA FISH
OT  - RNA localization
OT  - Sarcomere
OT  - Skeletal muscle
EDAT- 2024/03/19 18:42
MHDA- 2024/03/21 12:46
CRDT- 2024/03/19 12:21
PHST- 2024/03/21 12:46 [medline]
PHST- 2024/03/19 18:42 [pubmed]
PHST- 2024/03/19 12:21 [entrez]
AID - 10.1007/978-1-0716-3766-1_11 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2024;2784:163-176. doi: 10.1007/978-1-0716-3766-1_11.