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Methods Mol Biol. 2018;1649:127-142. doi: 10.1007/978-1-4939-7213-5_8.

Single mRNA Molecule Detection in Drosophila.

Author information

1
Department of Cell and Developmental Biology, University of Pennsylvania Perelman School of Medicine, 1112 Biomedical Research Bldg. II/III, 421 Curie Blvd., Philadelphia, PA, 19104, USA. shlittle@mail.med.upenn.edu.
2
Department of Physics, Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.

Abstract

Single molecule fluorescent in situ hybridization (smFISH) enables quantitative measurements of gene expression and mRNA localization. The technique is increasingly popular for analysis of cultured cells but is not widely applied to intact organisms. Here, we describe a method for labeling and detection of single mRNA molecules in whole embryos of the fruit fly Drosophila melanogaster. This method permits measurements of gene expression in absolute units, enabling new studies of transcriptional mechanisms underlying precision and reproducibility in cell specification.

KEYWORDS:

Drosophila; FISH; Fluorescence imaging; Quantitation; Single molecule

PMID:
29130194
PMCID:
PMC5772605
DOI:
10.1007/978-1-4939-7213-5_8
[Indexed for MEDLINE]
Free PMC Article

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