Application of phase-retrieved pupil functions in wide-field fluorescence microscopy

Bridget M. Hanser; Mats G. L. Gustafsson; David A. Agard; John W. Sedat

doi:10.1117/12.467844

15 May 2002 Application of phase-retrieved pupil functions in wide-field fluorescence microscopy

Bridget M. Hanser, Mats G. L. Gustafsson, David A. Agard, John W. Sedat

Proceedings Volume 4621, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX; (2002) https://doi.org/10.1117/12.467844
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

Multidimensional fluorescence microscopy data are often collected under imaging conditions that cause aberrations. For proper deconvolution of such data, one needs to generate a Point Spread Function (PSF) that includes the aberrations. Even if it is possible to model the imaging aberrations theoretically, there remains the problem of modifying the PSF accordingly. Measured PSFs are difficult to modify, and theoretical PSFs lack lens- and system-specific features. Phase retrieval approaches can be used to produce a compact and modifiable description of a microscope system, including its measured, system-specific performance. These pupil functions can be used in a depth-dependent deconvolution to correct for geometric distortions and improve restoration of relative intensities compared to spatially invariant deconvolution using unaberrated PSFs.

Citation Download Citation

Bridget M. Hanser, Mats G. L. Gustafsson, David A. Agard, and John W. Sedat "Application of phase-retrieved pupil functions in wide-field fluorescence microscopy", Proc. SPIE 4621, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX, (15 May 2002); https://doi.org/10.1117/12.467844

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