A Parallel Product-Convolution approach for representing the depth varying Point Spread Functions in 3D widefield microscopy based on principal component analysis.

We address the problem of computational representation of image formation in 3D widefield fluorescence microscopy with depth varying spherical aberrations. We first represent 3D depth-dependent point spread functions (PSFs) as a weighted sum of basis functions that are obtained by principal component analysis (PCA) of experimental data. This representation is then used to derive an approximating structure that compactly expresses the depth variant response as a sum of few depth invariant convolutions pre-multiplied by a set of 1D depth functions, where the convolving functions are the PCA-derived basis functions. The model offers an efficient and convenient trade-off between complexity and accuracy. For a given number of approximating PSFs, the proposed method results in a much better accuracy than the strata based approximation scheme that is currently used in the literature. In addition to yielding better accuracy, the proposed methods automatically eliminate the noise in the measured PSFs.

[1]  S. Gibson,et al.  Experimental test of an analytical model of aberration in an oil-immersion objective lens used in three-dimensional light microscopy. , 1992, Journal of the Optical Society of America. A, Optics and image science.

[2]  A. Nehorai,et al.  Deconvolution methods for 3-D fluorescence microscopy images , 2006, IEEE Signal Processing Magazine.

[3]  Taco D. Visser,et al.  Comparison of different theories for focusing through a plane interface , 1997 .

[4]  M. Gustafsson,et al.  Phase‐retrieved pupil functions in wide‐field fluorescence microscopy , 2004, Journal of microscopy.

[5]  Franck Marchis,et al.  AIDA: an adaptive image deconvolution algorithm with application to multi-frame and three-dimensional data. , 2007, Journal of the Optical Society of America. A, Optics, image science, and vision.

[6]  Joshua W Shaevitz,et al.  Enhanced three-dimensional deconvolution microscopy using a measured depth-varying point-spread function. , 2007, Journal of the Optical Society of America. A, Optics, image science, and vision.

[7]  Chrysanthe Preza,et al.  Image estimation accounting for point-spread function depth cariation in three-dimensional fluorescence microscopy , 2003, SPIE BiOS.

[8]  D. Agard,et al.  Fluorescence microscopy in three dimensions. , 1989, Methods in cell biology.

[9]  Michael Unser,et al.  A Fast Multilevel Algorithm for Wavelet-Regularized Image Restoration , 2009, IEEE Transactions on Image Processing.

[10]  S. Gibson,et al.  Experimental test of an analytical model of aberration in an oil-immersion objective lens used in three-dimensional light microscopy. , 1991, Journal of the Optical Society of America. A, Optics and image science.

[11]  Chrysanthe Preza,et al.  Depth-variant maximum-likelihood restoration for three-dimensional fluorescence microscopy. , 2004, Journal of the Optical Society of America. A, Optics, image science, and vision.

[12]  Michael Unser,et al.  A Fast Thresholded Landweber Algorithm for Wavelet-Regularized Multidimensional Deconvolution , 2008, IEEE Transactions on Image Processing.