Image analysis – quantitative in situ hybridization (qISH)
Image analysis is performed using Visiopharm’s combined stereology and supervised pixel classifier applications (color segmentation). This allows blinded quantitation of different parameters reflecting the presence of expression such as the stained areas, staining intensities or the frequency or average numbers of positive cells. This set-up is ideal for prognostic, diagnostic and drug response studies in larger cohorts.
Quantitation of in situ hybridization signal
Semiquantitative estimates of microRNA expression can be obtained by combining image sampling based on stereological principles with image analysis based on pixel classification. In this figure, the blue chromogenic ISH signal (panel A) is converted into green color (panel B), whereas the nuclear red stain is converted into red. Various expression parameters can be obtained from the classified images, such as areas and intensities.