Visualization of individual Scr mRNAs during Drosophila embryogenesis yields evidence for transcriptional bursting.

ABSTRACT: The detection and counting of transcripts within single cells via fluorescent in situ hybridization (FISH) has allowed researchers to ask quantitative questions about gene expression at the level of individual cells. This method is often preferable to quantitative RT-PCR, because it does not necessitate destruction of the cells being probed and maintains spatial information that may be of interest. Until now, studies using FISH at single-molecule resolution have only been rigorously carried out in isolated cells (e.g., yeast cells or mammalian cell culture). Here, we describe the detection and counting of transcripts within single cells of fixed, whole-mount Drosophila embryos via a combination of FISH, immunohistochemistry, and image segmentation. Our method takes advantage of inexpensive

SUBMITTER: Pare A 

PROVIDER: S-EPMC2805773 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature