Assessing the reliability of the suppose enhanced deconvolution method applied to dierent uorescent imaging techniques

TOSCANI, MICAELA; LACAPMESURE, AXEL M.; BRINATTI VAZQUEZ, GUILLERMO D.; MARTINEZ, OSCAR E.; MARTÍNEZ, SANDRA RITA

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Encuentro; 65 Annual Meeting of the Biophysical Society; 2021

The SUPPOSe algorithm converts the ill posed deconvolution problem to awell posed one using the ansatz that the object can be approximated by a superposition of virtual point sources of equal intensities. This source distribution isoptimized by comparing the measured image with an approximate image that isobtained by convolving the point spread function with the source distribution.As no regularization term is added a threefold to fivefold improvement in theresolution was shown but the improvement factor depends strongly on the noiseof the image and the sparsity of the sources. Moreover the genetic algorithmused for the optimization process and the noise could derive in artifacts inthe reconstruction. In this work we apply SUPPOSe under different contextsto assess the reliability of the retrieval by SUPPOSe. One scenario explored106a Wednesday, February 24, 2021is to perform different acquisitions of the same object to determine the precision of the reconstruction (i.e. fluctuations or changes under different realizations of the noise). To show the repeatability of the reconstructed images weused consecutive acquisitions of fluorescent microscopy images of F-actin filaments and microtubules over the same region after refocusing. The differencesbetween images due to both the different noise realization and the randomnessof the genetic algorithm are quantified by introducing a metric computation ofthe distance between solutions. In a second study, the accuracy of the methodunder different noise levels is discussed by applying a similar metric for thecomparison of the retrieved solution with the ground truth of synthetic images.Two photon microscopy images and also the improvement of STED images ofthe nuclear pores complex will also be presented to discuss the accuracy of thereconstruction.