Biomedical subjects

J A Conchello

Publications and source records attributed to J A Conchello.

4 recordsLinked to original sources

PubMed1999

Theoretical development and experimental evaluation of imaging models for differential-interference-contrast microscopy.

Imaging models for differential-interference-contrast (DIC) microscopy are presented. Two- and three-dimensional models for DIC imaging under partially coherent illumination were derived and tested by using phantom specimens viewed with several conventional DIC microscopes and quasi-monochromatic light. DIC images recorded with a CCD camera were compared with model predictions that were generated by using theoretical point-spread functions, computer-generated phantoms, and estimated imaging parameters such as bias and shear. Results show quantitative and qualitative agreement between model and data for several imaging conditions.

Evaluation Studies as Topic↗

PubMed1999

Parametric blind deconvolution: a robust method for the simultaneous estimation of image and blur.

Blind-deconvolution microscopy, the simultaneous estimation of the specimen function and the point-spread function (PSF) of the microscope, is an underdetermined problem with nonunique solutions that are usually avoided by enforcing constraints on the specimen function and the PSF. We derived a maximum-likelihood-based method for blind deconvolution in which we assume a mathematical model for the PSF that depends on a small number of parameters (e.g., less than 20). The algorithm then estimates the unknown parameters together with the specimen function. The mathematical model ensures that all the constraints of the PSF are satisfied, and the maximum-likelihood approach ensures that the specimen is nonnegative. The method successfully estimates the PSF and removes out-of-focus blur. The PSF estimation is robust to aberrations in the PSF and to noise in the image.

Algorithms↗

PubMed1998

Superresolution and convergence properties of the expectation-maximization algorithm for maximum-likelihood deconvolution of incoherent images.

Computational optical-sectioning microscopy with a nonconfocal microscope is fundamentally limited because the optical transfer function, the Fourier transform of the point-spread function, is exactly zero over a conic region of the spatial-frequency domain. Because of this missing cone of optical information, images are potentially artifactual. To overcome this limitation, superresolution, in the sense of band extrapolation, is necessary. I present a frequency-domain analysis of the expectation-maximization algorithm for maximum-likelihood image estimation that shows how the algorithm achieves this band extrapolation. This analysis gives the theoretical absolute bandwidth of the restored image; however, this absolute value may not be realistic in many cases. Then a second analysis is presented that assumes a Gaussian point-spread function and a specimen function and shows more realistic behavior of the algorithm and demonstrates some of its properties. Experimental results on the superresolving capability of the algorithm are also presented.

Algorithms↗

PubMed1994

Artifacts in computational optical-sectioning microscopy.

We tested the most complete optical model available for computational optical-sectioning microscopy and obtained four main results. First, we observed good agreement between experimental and theoretical point-spread functions (PSF's) under a variety of imaging conditions. Second, using these PSF's, we found that a linear restoration method yielded reconstructed images of a well-defined phantom object (a 10-microns-diameter fluorescent bead) that closely resembled the theoretically determined, best-possible linear reconstruction of the object. Third, this best linear reconstruction suffered from a (to our knowledge) previously undescribed artifactual axial elongation whose principal cause was not increased axial blur but rather the conical shape of the null space intrinsic to nonconfocal three-dimensional (3D) microscopy. Fourth, when 10-microns phantom beads were embedded at different depths in a transparent medium, reconstructed bead images were progressively degraded with depth unless they were reconstructed with use of a PSF determined at the bead's depth. We conclude that (1) the optical model for optical sectioning is reasonably accurate; (2) if PSF shift variance cannot be avoided by adjustment of the optics, then reconstruction methods must be modified to account for this effect; and (3) alternative microscopical or nonlinear algorithmic approaches are required for overcoming artifacts imposed by the missing cone of frequencies that is intrinsic to nonconfocal 3D microscopy.

Artifacts↗