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.

Relationships

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

Download All Files (To download individual files, select them in the “Files” panel above)

Download will include text file of metadata.



Globus is for large data sets.   What is Globus?

Analytics

Analytics