Computational optical sectioning microscopy (COSM) is gaining acceptance as an alternative as well as a complement to confocal scanning microscopes (CSM) and two-photo fluorescence excitation microscopes (TPFEM). COSM algorithms remove out-of-focus light from wide-field microscope images and can also improve the resolution in CSMs and TPFEMs. The algorithms require knowledge of the process of image formation usually represented by the point-spread function (PSF), the image of a point source of light. In modern scanning microscopes, the measurement of computation of the PSF is difficult at best. In these cases, it is necessary to estimate the PSF and the specimen fluorescence distribution simultaneously from the image; an approach called blind deconvolution (BD). We developed a method for blind deconvolution in which the PSF is constrained to follow a mathematical model that depends on a small number of unknown parameters. The parametric BD (PBD) method determines these parameters together with the specimen function. We have found encouraging results but the PBD is still computationally intensive. The purpose of this research is to9 make this method practical for routine use and to extend its application to CSMs and TOFEMS. The method will be optimized and its capabilities and limitations thoroughly assessed to be able to provide guidelines for its use. In addition, the method will be incorporated into a deconvolution package that our group developed.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM049798-08
Application #
6180213
Study Section
Special Emphasis Panel (ZRG1-SSS-I (02))
Program Officer
Deatherage, James F
Project Start
1993-08-01
Project End
2002-07-31
Budget Start
2000-08-01
Budget End
2001-07-31
Support Year
8
Fiscal Year
2000
Total Cost
$222,551
Indirect Cost
Name
Washington University
Department
Type
Other Domestic Higher Education
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Conchello, Jose-Angel; Dresser, Michael E (2007) Extended depth-of-focus microscopy via constrained deconvolution. J Biomed Opt 12:064026
Preza, Chrysanthe; Conchello, Jose-Angel (2004) Depth-variant maximum-likelihood restoration for three-dimensional fluorescence microscopy. J Opt Soc Am A Opt Image Sci Vis 21:1593-601
Markham, Joanne; Conchello, Jose-Angel (2003) Numerical evaluation of Hankel transforms for oscillating functions. J Opt Soc Am A Opt Image Sci Vis 20:621-30
Markham, J; Conchello, J A (2001) Fast maximum-likelihood image-restoration algorithms for three-dimensional fluorescence microscopy. J Opt Soc Am A Opt Image Sci Vis 18:1062-71
Markham, J; Conchello, J A (2001) Artefacts in restored images due to intensity loss in three-dimensional fluorescence microscopy. J Microsc 204:93-8
Preza, C; Snyder, D L; Conchello, J A (1999) Theoretical development and experimental evaluation of imaging models for differential-interference-contrast microscopy. J Opt Soc Am A Opt Image Sci Vis 16:2185-99
Markham, J; Conchello, J A (1999) Parametric blind deconvolution: a robust method for the simultaneous estimation of image and blur. J Opt Soc Am A Opt Image Sci Vis 16:2377-91
Conchello, J A (1998) Superresolution and convergence properties of the expectation-maximization algorithm for maximum-likelihood deconvolution of incoherent images. J Opt Soc Am A Opt Image Sci Vis 15:2609-19