Work Description

Title: Dataset of live-cell movies of single PolC-PAmCherry molecules in Bacillus subtilis cells with high and low fluorescent backgrounds. Open Access Deposited

http://creativecommons.org/licenses/by-nc/4.0/
Attribute Value
Methodology
  • We imaged Bacillus subtilis strains natively expressing the DNA polymerase PolC fused to the photoactivatable fluorescent protein PAmCherry as the sole source of PolC (strain JWS213). To produce the high background, a constant 15 W/cm2, 488-nm laser illumination generated a strong autofluorescent background in the cells; this background was further complicated by its slow decay over time. By stochastically switching a small subset (1 – 3 molecules per cell) of the PolC-PAmCherry molecules into a fluorescent state at a time (in a single-particle tracking/PALM experiment), we visualized the dynamics of single PolC-PAmCherry molecules in high-background cells and single PolC-PAmCherry molecules in low-background cells.
Description
  • This is the experimental data referenced in our manuscript entitled “SMALL-LABS: An algorithm for measuring single molecule intensity and position in the presence of obscuring backgrounds .” These live-cell single-molecule imaging movies were used as a test of the SMALL-LABS single-molecule image analysis algorithm. The dataset comprises two movies; each one is provided both as a .tif stack and as an .avi file. The movie called “low_bg” has a standard low background, and the movie called “high_bg” includes a high fluorescent background produced by an external 488-nm laser.
Creator
Depositor
  • isaacoff@umich.edu
Contact information
Discipline
Funding agency
  • National Science Foundation (NSF)
ORSP grant number
  • F050316-094587
Keyword
Citations to related material
  • B.P. Isaacoff, Y. Li, S.A. Lee, J.S. Biteen, "SMALL-LABS: Measuring Single-Molecule Intensity and Position in Obscuring Backgrounds." Biophysical Journal, 975-982, 116, 2019. DOI: 10.1016/j.bpj.2019.02.006
Resource type
Last modified
  • 03/21/2019
DOI
License
To Cite this Work:
Isaacoff, B., Li, Y., Lee, S., Biteen, J. Dataset of live-cell movies of single PolC-PAmCherry molecules in Bacillus subtilis cells with high and low fluorescent backgrounds [Data set]. University of Michigan - Deep Blue.

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Files (Count: 5; Size: 42.6 MB)

Thumbnail Title Original Upload Last Modified File Size Access Actions
Rf55z850j?file=thumbnail DeepBlueReadme.pdf 2018-09-28 2018-09-28 63 KB Open Access
Pz50gw96r?file=thumbnail high_bg.tif 2018-09-10 2018-09-10 17.8 MB Open Access
Default low_bg.avi 2018-09-10 2018-09-10 5.28 MB Open Access
5x21tg32p?file=thumbnail low_bg.tif 2018-09-10 2018-09-10 10.7 MB Open Access
Default high_bg.avi 2018-09-10 2018-09-10 8.8 MB Open Access

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