: Microscopy and, in particular, fluorescence microscopy plays an ever increasing role as a tool to improve the understanding of cellular processes such as receptor clustering and trafficking. The present application originated from a number of immunological studies which are being carried out by the co-PI. These studies relate to questions of recognition by the T cell receptor in autoimmune disease models (experimental autoimmune encephalomyelitis (EAE); collagen induced arthritis (CIA) and trafficking of the MHC class I-related receptor, FcRn. The overall objective of this multidisciplinary bioengineering project is to provide important additional tools for the analysis of experimental results obtained using fluorescence microscopy, with the emphasis on tools for three dimensional image sets. The premise of the application is that in a number of respects the analysis capabilities lag behind the recent developments in hardware and sample preparation methodology. The proposed study will investigate new methods and approaches to various problems in immunology and cellular biology. All proposed approaches will be rigorously analyzed with simulated and experimental data.
Our specific aims are: first, to investigate methods for quantitative analysis of clustering and co-localization. Second, deconvolution algorithms and several modifications will be analyzed. Third, a software suite will be written for acquisition and analysis of fluorescence microscopy images. The capabilities of this package will not only include the necessary functionality to carry out the earlier specific aims, but will provide a powerful development environment for advanced image analysis for microscopy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI050747-02
Application #
6620579
Study Section
Special Emphasis Panel (ZRG1-SSS-4 (02))
Program Officer
Johnson, David R
Project Start
2002-03-01
Project End
2005-02-28
Budget Start
2003-03-01
Budget End
2004-02-29
Support Year
2
Fiscal Year
2003
Total Cost
$262,930
Indirect Cost
Name
University of Texas-Dallas
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
800188161
City
Richardson
State
TX
Country
United States
Zip Code
75080
Chao, Jerry; Ward, E Sally; Ober, Raimund J (2010) A software framework for the analysis of complex microscopy image data. IEEE Trans Inf Technol Biomed 14:1075-87
Ram, Sripad; Ward, E Sally; Ober, Raimund J (2006) Beyond Rayleigh's criterion: a resolution measure with application to single-molecule microscopy. Proc Natl Acad Sci U S A 103:4457-62
Lai, X; Lin, Zhiping; Ward, E S et al. (2005) Noise suppression of point spread functions and its influence on deconvolution of three-dimensional fluorescence microscopy image sets. J Microsc 217:93-108
Vaccaro, Carlos; Zhou, Jinchun; Ober, Raimund J et al. (2005) Engineering the Fc region of immunoglobulin G to modulate in vivo antibody levels. Nat Biotechnol 23:1283-8
Ward, E Sally; Martinez, Cruz; Vaccaro, Carlos et al. (2005) From sorting endosomes to exocytosis: association of Rab4 and Rab11 GTPases with the Fc receptor, FcRn, during recycling. Mol Biol Cell 16:2028-38
Prabhat, Prashant; Ram, Sripad; Ward, E Sally et al. (2004) Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions. IEEE Trans Nanobioscience 3:237-42
Ober, Raimund J; Martinez, Cruz; Lai, Xuming et al. (2004) Exocytosis of IgG as mediated by the receptor, FcRn: an analysis at the single-molecule level. Proc Natl Acad Sci U S A 101:11076-81
Ober, Raimund J; Ram, Sripad; Ward, E Sally (2004) Localization accuracy in single-molecule microscopy. Biophys J 86:1185-200
Ward, E Sally; Zhou, Jinchun; Ghetie, Victor et al. (2003) Evidence to support the cellular mechanism involved in serum IgG homeostasis in humans. Int Immunol 15:187-95