New automatized approach for image processing in Xenopus laevis oocytes

GIBSON, OLIVER M.; GUASTAFERRI, FLORENCIA V.; CAVIGLIA, AGUSTÍN F.; AMODEO, GABRIELA; OZU, MARCELO; GALIZIA, LUCIANO

Congreso; Primeras Jornadas Virtuales SAB 2020; 2020

Sociedad Argentina de Biofísica

The Xenopus laevis oocyte is a cellular model used widely for aquaporin and ion channel heterologous expression. This cellular model allows recording sequential images or videos with low-cost videomicroscopy equipment. Our group developed different acquisition devices to study various aspects of the aquaporin functionality. Although this model would potentially allow obtaining data from thousands of oocytes from each donor frog, one bottleneck in the experimental workflow is image processing. Timelapses recorded at high temporal resolution (e.g., 30 fps), may contain thousands of images from which cell volume information must be extracted. The traditional processing of those images is performed manually, using free software (e.g., ImageJ) to segment the oocyte and to evaluate cell volume under high circularity criteria. Oocyte segmentation is user-dependent and requires training and experimental criteria. Then, information about the oocyte area in each image is converted to a relative volume curve. The water osmotic permeability coefficient (Pf) is then calculated using the slope of the Vt/V0 curve. These sequential steps are time-consuming. The challenge of this work is to present a new Phyton-based tool which allows processing images, extract the information and calculate Vt/V0 curves and the  Pf of each oocyte. We developed this new script to produce an automatic, faster, and operator-independent analysis of Pf. The results are compared with the manual/traditional approach.