Imaging individual mRNA molecules using multiple singly labeled probes.
We describe a method for imaging individual mRNA molecules in fixed cells by probing each mRNA species with 48 or more short, singly labeled oligonucleotide probes. This makes each mRNA molecule visible as a computationally identifiable fluorescent spot by fluorescence microscopy. We demonstrate simultaneous detection of three mRNA species in single cells and mRNA detection in yeast, nematodes, fruit fly wing discs, and mammalian cell lines and neurons.
Raj
Arjun
A
Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. arjunraj@mit.edu
van den Bogaard
Patrick
P
Rifkin
Scott A
SA
van Oudenaarden
Alexander
A
Tyagi
Sanjay
S
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D008322:Mammals; D015335:Molecular Probes; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics
D015335:Molecular Probes; D012333:RNA, Messenger