title: Single molecule imaging of RNA in situ.
abstract: This protocol describes a method to image individual mRNA molecules in situ. About 50 oligonucleotides complementary to different regions of a target mRNA species are used simultaneously. Each probe is labeled with a single fluorescent moiety. When these probes bind to their target, each mRNA molecule becomes so intensely fluorescent that it can be seen as a fine fluorescent spot. Several different mRNA species can be detected in multiplex imaging using differently colored probe sets for each species. An automated image-processing program is used to count the number of mRNA molecules of each species that are expressed in each cell, thus yielding single-cell gene expression profiles.
authors: Public Health Research Institute Center, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, NJ, USA.
authors: Mona
authors: M
authors: Batish
authors: Arjun
authors: A
authors: Raj
authors: Sanjay
authors: S
authors: Tyagi
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D057054:Molecular Imaging / Q000379:methods*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012313:RNA / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051381:Rats
chemical_list: D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D012313:RNA