Abstract: This research analyses Discrete Cosine Transform based Gradient Vector Flow Active Contours, a segmentation tool for chromosome spread images, which have hitherto been established as an efficient segmentation tool. Extensive research has been performed on Discrete Cosine Transform based Gradient Vector Flow Active Contours for segmentation of chromosome spread images. This study is an extended analysis in the same area. This study yields more insight on efficient segmentation of chromosome spread images using Discrete Cosine Transform based Gradient Vector Flow Active Contours.