Characterization of a 3D microscope imaging system

Steven S. S. Poon; Stephen J. Lockett; Rabab K. Ward

doi:10.1117/12.148626

29 July 1993 Characterization of a 3D microscope imaging system

Steven S. S. Poon, Stephen J. Lockett, Rabab K. Ward

Proceedings Volume 1905, Biomedical Image Processing and Biomedical Visualization; (1993) https://doi.org/10.1117/12.148626
Event: IS&T/SPIE's Symposium on Electronic Imaging: Science and Technology, 1993, San Jose, CA, United States

Abstract

To reconstruct the object from its observed images, the characteristics of the imaging system must first be obtained. In a microscope imaging system, the characteristics vary not only in the imaging plane, but also vary as a function of the focus in which the image is taken. Thus, a three dimensional system response or point spread function (PSF) needs to be determined. One way of determining the PSF is to use a theoretical approach to analyze the aberration-free microscope imaging system. However, the assumptions and properties of lenses in the system are often not ideal. Thus an experimental approach for determining the PSF is sometimes used. We report on the results of our experiments where some of the problems associated with the determination of the experimental PSF are overcome. Point source objects are hard to find in nature. In our analysis, we use test objects which simulates point sources, (such as small fluorescence beads,) and objects which can be described as a convolution of the PSF with their shape (such as a step edge). To increase the spatial resolution, the precise location of the object is also estimated to a fraction of a pixel. The results of this is then compared with the theoretical approach.

Citation Download Citation

Steven S. S. Poon, Stephen J. Lockett, and Rabab K. Ward "Characterization of a 3D microscope imaging system", Proc. SPIE 1905, Biomedical Image Processing and Biomedical Visualization, (29 July 1993); https://doi.org/10.1117/12.148626

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