Automatic Analysis ofTUNELassay Microscopy Images

Acomputer methodfor theautomatic analysis ofspecific collected usinga cooled CCD (charged coupled device) fluorescence microscopy images, collected witha TUNELassaycameraandcaptured byusingQuips®PathVysion (Smart procedure applied onhumanspermatozoa cells, hasbeendeveloped Capture VP)anddualbandpassfilter (DAPI/Orange). inthis work. Ineachimage, thecells areshowninblue color while intheinterior ofevery cell anareaproportional tothedegree ofthe cell DNAfragmentation hasbeenstained inreddish color bythe assay procedure. Theproposed digital imageanalysis technique suggests asegmentation method based onthePrimary Component Analysis (PCA)applied tothe3-Dcolor space. Inthis waythe shapes ineveryimageareisolated andthena classification technique isusedtoobtain onlytheshapes representing solitary spermatozoa, toignore those presenting celloverlapping and dispose smallshapes-scraps belonging tofragments ofuseless material created during theassay procedure. Finally, foreach solitary cell theratio ofthereddish areaoverthetotal cell areais estimated. ., ~~~~~~fragmented regions inlight graycolor. Thebackground, although itseems endlabelling (TUNEL)assaywasusedl withproper process uniform, presents fluctuations intheintensity which arenotvisible inthe and controls foreachslide(1).A Zeiss-Axioscope grayscale images! fluorescence microscope wasusedandtheimageswere