DOI: 10.1083/jcb.201411032. Nuclear accessibility of β-actin mRNA is measured by 3D single-molecule real-time tracking

Nuclear accessibility of β-actin mRNA is measured by 3D single-molecule real-time tracking

10.1083/jcb.201411032

Crossref journal-article

Rockefeller University Press

Journal of Cell Biology (291)

Abstract

Imaging single proteins or RNAs allows direct visualization of the inner workings of the cell. Typically, three-dimensional (3D) images are acquired by sequentially capturing a series of 2D sections. The time required to step through the sample often impedes imaging of large numbers of rapidly moving molecules. Here we applied multifocus microscopy (MFM) to instantaneously capture 3D single-molecule real-time images in live cells, visualizing cell nuclei at 10 volumes per second. We developed image analysis techniques to analyze messenger RNA (mRNA) diffusion in the entire volume of the nucleus. Combining MFM with precise registration between fluorescently labeled mRNA, nuclear pore complexes, and chromatin, we obtained globally optimal image alignment within 80-nm precision using transformation models. We show that β-actin mRNAs freely access the entire nucleus and fewer than 60% of mRNAs are more than 0.5 µm away from a nuclear pore, and we do so for the first time accounting for spatial inhomogeneity of nuclear organization.

Bibliography

Smith, C. S., Preibisch, S., Joseph, A., Abrahamsson, S., Rieger, B., Myers, E., Singer, R. H., & Grunwald, D. (2015). Nuclear accessibility of Î²-actin mRNA is measured by 3D single-molecule real-time tracking. Journal of Cell Biology, 209(4), 609â619.

Authors 8

Carlas S. Smith (first)
Stephan Preibisch (additional)
Aviva Joseph (additional)
Sara Abrahamsson (additional)
Bernd Rieger (additional)
Eugene Myers (additional)
Robert H. Singer (additional)
David Grunwald (additional)

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Dates

Type	When
Created	10 years, 2 months ago (May 25, 2015, 2:18 p.m.)
Deposited	2 years, 1 month ago (July 23, 2023, 3:27 a.m.)
Indexed	1 month ago (July 20, 2025, 12:07 a.m.)
Issued	10 years, 2 months ago (May 25, 2015)
Published	10 years, 2 months ago (May 25, 2015)
Published Online	10 years, 2 months ago (May 25, 2015)
Published Print	10 years, 2 months ago (May 25, 2015)

Funders 0

None

BibTeX

@article{Smith_2015, title={Nuclear accessibility of β-actin mRNA is measured by 3D single-molecule real-time tracking}, volume={209}, ISSN={0021-9525}, url={http://dx.doi.org/10.1083/jcb.201411032}, DOI={10.1083/jcb.201411032}, number={4}, journal={Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Smith, Carlas S. and Preibisch, Stephan and Joseph, Aviva and Abrahamsson, Sara and Rieger, Bernd and Myers, Eugene and Singer, Robert H. and Grunwald, David}, year={2015}, month=may, pages={609–619} }

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