AI Article Synopsis
- Live-cell single molecule imaging has advanced our understanding of how DNA-binding molecules like transcription factors navigate the nucleus to find their target sites.
- Traditional single molecule experiments have been restricted to thin cell cultures, but lattice light-sheet microscopy now allows for imaging in thicker specimens like embryos.
- This text outlines a protocol for imaging transcription factor dynamics in Drosophila melanogaster embryos and suggests its applicability to other organisms, including live mouse embryos.
Article 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.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6225527 | PMC |
| http://dx.doi.org/10.1007/978-1-4939-8591-3_32 | DOI Listing |
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