Toward the specificity in QPI 3D Tomographic Cell Flow Cytometry Holography: recent achievements and perspectives in biomedical sciences

The actual gap of the label-free quantitative phase microscopy in respect to fluorescence microscopy, that allows the subcellular characterization by using exogenous markers, is the lack of intracellular specificity. Recently, computational methods based on artificial intelligence have been demonstrated, which allow a virtual staining of single cells in both 2D and 3D, but they require co-registration systems able to collect simultaneously both fluorescence and quantitative phase information. However, a real limitation exists, i.e. these approaches cannot be used in flow cytometry condition. In this paper, we discuss a new methodology for adding the intracellular specificity analysis to tomographic phase microscopy in flow cytometry. The proposed strategy is based on the statistical clustering of tomograms voxels, thus allowing the segmentation of cell’s organelles. Here we report the results of nuclear region identification for cancer cells.