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

Attribute Value
  • 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.
  • 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.
Contact information
Funding agency
  • National Science Foundation (NSF)
Other funding agency
  • National Science Foundation (NSF)
ORSP grant number
  • F050316-094587
  • B.P. Isaacoff, Y. Li, S.A. Lee, and J.S. Biteen, “SMALL-LABS: An algorithm for measuring single molecule intensity and position in the presence of obscuring backgrounds,” submitted to Biophysical Journal on April 18, 2018.
Resource type
Last modified
  • 09/28/2018
  • doi:10.7302/Z2CR5RKD
CC License


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