Quantitative spatial analysis of transcripts in multinucleate cells using single-molecule FISH.

ABSTRACT: mRNA positioning in the cell is important for diverse cellular functions and proper development of multicellular organisms. Single-molecule RNA FISH (smFISH) enables quantitative investigation of mRNA localization and abundance at the level of individual molecules in the context of cellular features. Details about spatial mRNA patterning at various times, in different genetic backgrounds, at different developmental stages, and under varied environmental conditions provide invaluable insights into the mechanisms and functions of spatial regulation. Here, we describe detailed methods for performing smFISH along with immunofluorescence for two large, multinucleate cell types: the fungus Ashbya gossypii and cultured mouse myotubes. We also put forward a semi-automated image processing tool tha

SUBMITTER: Lee C 

PROVIDER: S-EPMC4808427 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature