In collaboration with the laboratory of Dr, Flemming Hansen, Technical University of Denmark, we have developed a fully automated cell recognition and fluorescent focus-measuring program in the form of a macro within the image analysis program Image Pro Plus. This has been very successful in the basic tasks of detecting fluorescent foci formed by plasmids and specific chromosomal loci in cells of the bacterium Escherichia coli. As our needs for analysis of fluorescent images become more demanding, we continue to develop and adapt this system to meet them. We have developed an analytical tool """"""""Inspect"""""""" that facilitates the analysis of images that have two color fluorescence information. In conjunction with the data collection programs, we have developed an array of spreadsheet programs to analyze and portray the data. We have completed a major new thrust in the development of the image analysis software. This allows the automated identification of multiple classes of cells in various states of division, and of foci that are dividing etc. These cells can then be inspected directly as a sub-class of images extracted for inspection. New sub-classes can then be defined and analyzed with seamless switching between the numerical data and the individual cell images. This allows us to analyze large populations in far more detail than was previously possible. This year we developed extensive software to handle the extended data collecting capability that we developed in 2011.Special emphasis has been laid on the ability to analize the cell short axis data and to project distributions of markers to the radial axis. This allows us to predict. the dynamic form of the replicating and segregating chromosomes in 3D.
|Sengupta, Manjistha; Nielsen, Henrik Jorck; Youngren, Brenda et al. (2010) P1 plasmid segregation: accurate redistribution by dynamic plasmid pairing and separation. J Bacteriol 192:1175-83|