Ellipsoid localization microscopy infers the size and order of protein layers in Bacillus spore coats. Manetsberger, J.; Manton, J. D.; Erdelyi, M. J.; Lin, H.; Rees, D.; Christie, G.; and Rees, E. J. Biophysical Journal, 109(10):2058--2066, November, 2015.
Ellipsoid localization microscopy infers the size and order of protein layers in Bacillus spore coats [link]Paper  doi  abstract   bibtex   
Multilayered protein coats are crucial to the dormancy, robustness, and germination of bacterial spores. In Bacillus subtilis spores, the coat contains over 70 distinct proteins. Identifying which proteins reside in each layer may provide insight into their distinct functions. We present image analysis methods that determine the order and geometry of concentric protein layers by fitting a model description for a spheroidal fluorescent shell image to optical micrographs of spores incorporating fluorescent fusion proteins. The radius of a spherical protein shell can be determined with \textless10 nm error by fitting an equation to widefield fluorescence micrographs. Ellipsoidal shell axes can be fitted with comparable precision. The layer orders inferred for B. subtilis and B. megaterium are consistent with measurements in the literature. The aspect ratio of elongated spores and the tendency of some proteins to localize near their poles can be quantified, enabling measurement of structural anisotropy.
@article{manetsberger_ellipsoid_2015,
	title = {Ellipsoid localization microscopy infers the size and order of protein layers in {Bacillus} spore coats},
	volume = {109},
	issn = {0006-3495},
	url = {http://www.cell.com/article/S0006349515009935/abstract},
	doi = {10.1016/j.bpj.2015.09.023},
	abstract = {Multilayered protein coats are crucial to the dormancy, robustness, and germination of bacterial spores. In Bacillus subtilis spores, the coat contains over 70 distinct proteins. Identifying which proteins reside in each layer may provide insight into their distinct functions. We present image analysis methods that determine the order and geometry of concentric protein layers by fitting a model description for a spheroidal fluorescent shell image to optical micrographs of spores incorporating fluorescent fusion proteins. The radius of a spherical protein shell can be determined with {\textless}10 nm error by fitting an equation to widefield fluorescence micrographs. Ellipsoidal shell axes can be fitted with comparable precision. The layer orders inferred for B. subtilis and B. megaterium are consistent with measurements in the literature. The aspect ratio of elongated spores and the tendency of some proteins to localize near their poles can be quantified, enabling measurement of structural anisotropy.},
	language = {English},
	number = {10},
	urldate = {2015-11-20TZ},
	journal = {Biophysical Journal},
	author = {Manetsberger, Julia and Manton, James D. and Erdelyi, Miklos J. and Lin, Henry and Rees, David and Christie, Graham and Rees, Eric J.},
	month = nov,
	year = {2015},
	pages = {2058--2066}
}
Downloads: 0