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Open AccessJournal ArticleDOI

Single Molecule Imaging of Transcription Factor Binding to DNA in Live Mammalian Cells

J. Christof M. Gebhardt, +10 more
- 01 May 2013 - 
- Vol. 10, Iss: 5, pp 421-426
TLDR
Reflected light-sheet microscopy (RLSM), a fluorescence microscopy method allowing selective plane illumination throughout the nuclei of living mammalian cells, is developed and single-molecule measurements and statistical analysis revealed dynamic properties of transcription factors.
Abstract: 
Light-sheet microscopy using a laser beam reflected off a mirrored AFM cantilever provides high signal-to-background images suitable for high-speed quantitative single-molecule imaging of transcription factor binding to DNA in the nucleus of living mammalian cells.

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Citations
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Journal ArticleDOI

The Human Transcription Factors.

Samuel A. Lambert, +12 more
- 08 Feb 2018 - 
TL;DR: This review considers how TFs are identified and functionally characterized, principally through the lens of a catalog of over 1,600 likely human TFs and binding motifs for two-thirds of them, highlighting the importance of continued effort to understand TF-mediated gene regulation.
Journal ArticleDOI

Intrinsically disordered proteins in cellular signalling and regulation.

Peter E. Wright, +1 more
- 01 Jan 2015 - 
TL;DR: Experimental, computational and bioinformatic analyses combine to identify and characterize disordered regions of proteins, leading to a greater appreciation of their widespread roles in biological processes.
Journal ArticleDOI

Lattice Light Sheet Microscopy: Imaging Molecules to Embryos at High Spatiotemporal Resolution

Bi-Chang Chen, +27 more
- 24 Oct 2014 - 
TL;DR: A new microscope using ultrathin light sheets derived from two-dimensional optical lattices is developed, demonstrating the performance advantages of lattice light-sheet microscopy compared with previous techniques and highlighted phenomena that, when seen at increased spatiotemporal detail, may hint at previously unknown biological mechanisms.
Journal ArticleDOI

Single-Molecule Dynamics of Enhanceosome Assembly in Embryonic Stem Cells

Jiji Chen, +12 more
- 13 Mar 2014 - 
TL;DR: In vivo and in vitro single-molecule imaging, transcription factor (TF) mutagenesis, and ChIP-exo mapping are combined to determine how TFs dynamically search for and assemble on their cognate DNA target sites and find that enhanceosome assembly is hierarchically ordered.
Journal ArticleDOI

A guide to light-sheet fluorescence microscopy for multiscale imaging

Rory M. Power, +5 more
- 01 Apr 2017 - 
TL;DR: This work elucidate the key developments and define a simple set of underlying principles governing LSFM, which aim to clarify the decisions to be made for those who wish to develop and use bespoke light-sheet systems and to assist in identifying the best approaches to apply this powerful technique to myriad biological questions.
References
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Journal ArticleDOI

Precise nanometer localization analysis for individual fluorescent probes

Russell E. Thompson, +2 more
- 01 May 2002 - 
TL;DR: A localization algorithm motivated from least-squares fitting theory is constructed and tested both on image stacks of 30-nm fluorescent beads and on computer-generated images (Monte Carlo simulations), and results show good agreement with the derived precision equation.
Journal ArticleDOI

Optical sectioning deep inside live embryos by selective plane illumination microscopy

Jan Huisken, +4 more
- 13 Aug 2004 - 
TL;DR: In this article, a selective plane illumination microscopy (SPIM) was developed to generate multidimensional images of samples up to a few millimeters in size, which can be applied to visualize the embryogenesis of the relatively opaque Drosophila melanogaster in vivo.
Journal ArticleDOI

Role of the CLOCK Protein in the Mammalian Circadian Mechanism

Nicholas Gekakis, +21 more
- 05 Jun 1998 - 
TL;DR: CLOCK-BMAL1 heterodimers appear to drive the positive component of per transcriptional oscillations, which are thought to underlie circadian rhythmicity.
Journal ArticleDOI

HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein Analysis

Georgyi V. Los, +19 more
- 06 Jun 2008 - 
TL;DR: The utility of this modular protein tagging system for cellular imaging and protein immobilization is demonstrated by analyzing multiple molecular processes associated with NF-kappaB-mediated cellular physiology, including imaging of subcellular protein translocation and capture of protein--protein and protein--DNA complexes.
Journal ArticleDOI

A general method for the covalent labeling of fusion proteins with small molecules in vivo

Antje Keppler, +5 more
- 01 Jan 2003 - 
TL;DR: A general method for the covalent labeling of fusion proteins in vivo that complements existing methods for noncovalentlabeling of proteins and that may open up new ways of studying proteins in living cells is described.
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