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Space non-invariant point-spread function and its estimation in fluorescence microscopy

Praveen Pankajakshan 1 Laure Blanc-Féraud 1 Zvi Kam 2 Josiane Zerubia 1
1 ARIANA - Inverse problems in earth monitoring
CRISAM - Inria Sophia Antipolis - Méditerranée , Laboratoire I3S - SIS - Signal, Images et Systèmes
Abstract : In this research report, we recall briefly how the diffraction-limited nature of an optical microscope's objective, and the intrinsic noise can affect the observed images' resolution. A blind deconvolution algorithm can restore the lost frequencies beyond the diffraction limit. However, under other imaging conditions, the approximation of aberration-free imaging, is not applicable, and the phase aberrations of the emerging wavefront from a specimen immersion medium cannot be ignored any more. We show that an object's location and its original intensity distribution can be recovered by retrieving the refracted wavefront's phase from the observed intensity images. We demonstrate this by retrieving the point-spread function from an imaged microsphere. The noise and the influence of the microsphere size can be mitigated and sometimes completely removed from the observed images by using a maximum a posteriori estimate. However, due to the incoherent nature of the acquisition system, phase retrieval from the observed intensities will be possible only if the phase is constrained. We have used geometrical optics to model the phase of the refracted wavefront, and tested the algorithm on some simulated images.
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Contributor : Praveen Pankajakshan <>
Submitted on : Friday, December 18, 2009 - 8:21:54 PM
Last modification on : Thursday, October 15, 2020 - 2:42:03 PM
Long-term archiving on: : Thursday, September 23, 2010 - 11:33:44 AM


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  • HAL Id : inria-00438719, version 2



Praveen Pankajakshan, Laure Blanc-Féraud, Zvi Kam, Josiane Zerubia. Space non-invariant point-spread function and its estimation in fluorescence microscopy. [Research Report] RR-7157, INRIA. 2009, pp.54. ⟨inria-00438719v2⟩



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