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PSF identification applied to 3-D fluorescence microscopy quantification

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Abstract

3-D optical fluorescence microscopy becomes now an efficient tool for volumic investigation of living biological samples. However, acquired raw data suffer from different distortions. In order to carry out biological analysis, restoration of raw data by deconvolution is mandatory. The system identification is useful to obtain the knowledge of the actual system and to quantify the influence of experimental parameters. High order centered moments are used as PSF descriptors. Oil immersion index, numerical aperture and specimen thickness are critical parameters for data quality. Furthermore, PSF identification is helpful to precise the experimental protocol. Application to 3-D anthracycline distribution in breast cancer cells is presented.

© 2001 OSA/SPIE

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