The Dynamic In Situ Cytometry (DISC) is a new approach that combines advantages of both intravital imaging and flow cytometry analysis. With the help of the custom designed DISCit software, the DISC provides the opportunity to link cell behavior and phenotype in vivo and enable to perform cytometry in vivo and in real-time. In addition it enables you to readily display and analyze multiparametric imaging data. The DISCit software is freely available upon request.
Time-lapse movie processing for DISC analysis.
Following intravital imaging of GFP+ cells in the presence of fluorescent Fab fragments (e.g. in the spleen), time-lapse movies are subjected to 3D cell tracking and cell segmentation. Time-lapse movies can then be further processed by applying a mask corresponding to the segmented cell volume. GFP and Fab fluorescence are then automatically extracted for each cell at each time point. Click here to see the movie.