A Tool for Alignment and Averaging of Sparse Fluorescence Signals in Rod-Shaped Bacteria.
Fluorescence microscopy studies have shown that many proteins localize to highly specific subregions within bacterial cells. Analyzing the spatial distribution of low-abundance proteins within cells is highly challenging because information obtained from multiple cells needs to be combined to provide well-defined maps of protein locations. We present (to our knowledge) a novel tool for fast, automated, and user-impartial analysis of fluorescent protein distribution across the short axis of rod-shaped bacteria. To demonstrate the strength of our approach in extracting spatial distributions and visualizing dynamic intracellular processes, we analyzed sparse fluorescence signals from single-molecule time-lapse images of individual Escherichia coli cells. In principle, our tool can be used to provide information on the distribution of signal intensity across the short axis of any rod-shaped object.
Zernike Institute for Advanced Materials, University of Groningen, Groningen, the Netherlands.
Joris M H
JMH
Goudsmits
Zernike Institute for Advanced Materials, University of Groningen, Groningen, the Netherlands; School of Chemistry, University of Wollongong, Wollongong, New South Wales, Australia.
Antoine M
AM
van Oijen
Zernike Institute for Advanced Materials, University of Groningen, Groningen, the Netherlands; School of Chemistry, University of Wollongong, Wollongong, New South Wales, Australia. Electronic address: andrewr@uow.edu.au.
Andrew
A
Robinson
D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D021381:Protein Transport
D029968:Escherichia coli Proteins