Understanding the mechanisms underlying neurological disorders and offering fresh viewpoints and remedies, neuroscientists must bridge the gap between emerging technology and experimental research. We computed the cell migration ratio of confocal pictures of astrocytes and glioblastoma (GBM) in our recent study. The primary goal is to comprehend how astrocyte placement in the 3D cell culture model influences GBM migration. For image processing applications, there are numerous computational tools available. MATLAB provides an extensive library and the ability to create new, bespoke functions on an intuitive platform. Hence, results are obtained by using MATLAB functions. Our goal was to quantify the area that cells that are migrated within the observed region. Further processing on the dataset formed by color images is achieved by taking the luminance channel of these color images. As a result of this, the highest contrast can be obtained, and segmentation of the cells can be calculated robustly. After that migration route can be determined. Using the identical cell line methods, confocal pictures were collected on days 3, 5, 7, and 9 in two sets of experiment series. Our observations showed that GBM cells were consistently outside of a particular circle. For each image, we therefore counted the area that cells occupy outside of a certain circle. These figures were acquired in micrometer and pixel squares. Next, we used the following formula to determine the migration capacity as a parameter, which is expressed as a percentage: Area Outside the Circle / Area in Pixel Net x 100.