Data correction methods for 3D light microscopy

Chuo-Lung Wang; Nathan J. O'Connor; Michel Wang; James N. Turner; Timothy J. Holmes

doi:10.1117/12.310544

9 June 1998 Data correction methods for 3D light microscopy

Chuo-Lung Wang, Nathan J. O'Connor, Michel Wang, James N. Turner, Timothy J. Holmes

Proceedings Volume 3261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V; (1998) https://doi.org/10.1117/12.310544
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

Abstract

A cooled or video-rate CCD camera is often used to collect optical slices from many modalities of microscopy including widefield-fluorescence (WF) and transmitted-light brightfield (TBL). Raw optical slices collected from both of these types of cameras are contaminated by imperfect performance of the CCD camera. Optical slice image data must be corrected for the bias level and nonuniformity in the photometric response of CCD elements. Fluctuation of the exposure time in a cooled CCD camera needs to be calibrated as well. Bad pixels due to the severely low sensitivity of some of the CCD elements often occur and otherwise impair the conventional schemes of correction and calibration. An adaptive median filter has been introduced by us previously to treat these bad pixels. In this study a Wiener-type regularization scheme is proposed as robust, fast, and practical alternative treatment of these bad pixels. One advantage of using this scheme is that doing so provides a nicely modularized design of the computer algorithm. As such, the correction scheme is software readily integrated as a software component in a 3D microscopy system and may operate independently without affecting the design of other software components in the system.

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Chuo-Lung Wang, Nathan J. O'Connor, Michel Wang, James N. Turner, and Timothy J. Holmes "Data correction methods for 3D light microscopy", Proc. SPIE 3261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V, (9 June 1998); https://doi.org/10.1117/12.310544

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KEYWORDS

Digital filtering

Data corrections

CCD cameras

Microscopy

Charge-coupled devices

Calibration

Image filtering

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