Quantitative depth-variant imaging for fluorescence microscopy using the COSMOS software package

Chrysanthe Preza; Vimeetha Myneni

doi:10.1117/12.840688

24 February 2010 Quantitative depth-variant imaging for fluorescence microscopy using the COSMOS software package

Chrysanthe Preza, Vimeetha Myneni

Proceedings Volume 7570, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVII; 757003 (2010) https://doi.org/10.1117/12.840688
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

The previously developed depth-variant expectation maximization (DVEM) restoration algorithm for fluorescent microscopy [1] has been implemented as part of the computational optical sectioning microscopy open source (COSMOS) software package [2]. COSMOS facilitates the development and dissemination of the DVEM algorithm which addresses depth-variant imaging due to spherical aberrations. Physical parameters such as the imaging lens, the size of the object and its refractive index contribute to the amount of spherical aberrations present in the image. The algorithm is based on a depth-variant imaging model and it requires a small number of point-spread functions (PSFs) defined at different depths within the sample. An acceptable choice for the number of PSFs needed for the computation provides a tradeoff between accuracy of the result and computational complexity. In this paper we show results obtained from simulation studies in which the performance of the DVEM algorithm is investigated for different parameters that contribute to depth variability. Some of the DVEM algorithm results are compared to results obtained with a spaceinvariant (or deconvolution) method from the same simulated data. The main conclusion from these studies is that a small number of PSFs can be used for the estimation without a significant loss in algorithm performance rendering the algorithm suitable for practical use. Furthermore, DVEM offers improvements over deconvolution methods with a small increase in computational complexity.

Citation Download Citation

Chrysanthe Preza and Vimeetha Myneni "Quantitative depth-variant imaging for fluorescence microscopy using the COSMOS software package", Proc. SPIE 7570, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVII, 757003 (24 February 2010); https://doi.org/10.1117/12.840688

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