Single molecule fluorescent in situ hybridization (smFISH) of C. elegans worms and embryos
Name
van Oudenaarden_Single molecule.pdf
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2.44 MB
Format
Adobe PDF
Checksum (MD5)
ddc7016fb9e1700057286d08853419b0
Author(s)
Ji, Ni
van Oudenaarden, Alexander
Date Issued
December 2012
Journal
WormBook
Publisher
WormBook
Citation
Ji, Ni, and Alexander van Oudenaarden. “Single Molecule Fluorescent in Situ Hybridization (smFISH) of C. Elegans Worms and Embryos.” WormBook (2012): 1–16.
Version
Final published version
Abstract
In C. elegans, the expression pattern of a gene provides important clues to understanding its biological function. To accurately depict endogenous transcriptional activity, a highly sensitive method is required to measure transcript levels in the intact tissue across various developmental stages. Conventional RNA in situ hybridization methods using hapten- (biotin or digoxygenin) labeled RNA probes rely on antibody binding for visualization, and are thus only semi-quantitative at best (Raap et al. 1995; Levsky et al. 2003). Additionally, hapten-labeled probes are prone to diffuse localization (when conjugated with alkaline phosphatase), low sensitivity (when conjugated with fluorescent molecules), and non-specific probe binding. Here, we introduce a recently developed mRNA in situ hybridization method (Raj et al. 2008) that circumvents the above difficulties to give single molecule resolution of transcript detection.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Physics
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1895/wormbook.1.153.1
