| Literature DB >> 29956254 |
Mustafa Mir1, Armando Reimer2, Michael Stadler1, Astou Tangara1, Anders S Hansen1, Dirk Hockemeyer1, Michael B Eisen1,2,3,4, Hernan Garcia1,2,5, Xavier Darzacq6.
Abstract
In the past decade, live-cell single molecule imaging studies have provided unique insights on how DNA-binding molecules such as transcription factors explore the nuclear environment to search for and bind to their targets. However, due to technological limitations, single molecule experiments in living specimens have largely been limited to monolayer cell cultures. Lattice light-sheet microscopy overcomes these limitations and has now enabled single molecule imaging within thicker specimens such as embryos. Here we describe a general procedure to perform single molecule imaging in living Drosophila melanogaster embryos using lattice light-sheet microscopy. This protocol allows direct observation of both transcription factor diffusion and binding dynamics. Finally, we illustrate how this Drosophila protocol can be extended to other thick samples using single molecule imaging in live mouse embryos as an example.Entities:
Keywords: Drosophila melanogaster; Lattice light-sheet microscopy; Live embryo imaging; Selective plane illumination microscopy; Single molecule fluorescence; Single molecule imaging; Single molecule kinetics; Single particle tracking; Transcription factor dynamics
Mesh：
Year: 2018 PMID： 29956254 PMCID： PMC6225527 DOI： 10.1007/978-1-4939-8591-3_32
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745