Molecular Cytology

ABSTRACT The major goal of the Molecular Cytology Core is to deliver cutting-edge, specialized services in support of research projects at the Center. The methodologies employed by the Core allow for the detection in situ (in cells, tissue sections, organoids and whole mounts) of biomarkers (proteins, glycoproteins, lipids, nucleic acids) and molecular processess in normal and pathological conditions. The precise localization of the biomarkers is achieved by optical imaging of fixed and live samples. Several molecular markers (up to seven) can be vizualized simultanously or sequentially in the same sample by multiplex staining, using machinebased protocols. In collaboration with its users, the Core performs comprehensive antibody validations. The Core trains investigators in basic staining principles and assists them in performing manual experiments. The optical imaging modalities within the Core allow for wide field imaging and confocal imaging of tissue sections, thick and cleared tissue samples, cells, live embryos, tumors in live mice, etc. Super-resolution imaging is performed as well as confocal imaging in the IR range to visualize nanomaterials and drugs. With the newly acquired module for fluorescence correlation spectroscopy (FCS) imaging, researchers can study the dynamic behavior of biomolecules. The Core scans user slides with four digital slide scanners, one of which has confocal capabilities. The scanning generates large volumes of data and serves as a valuable resource for image analysis. Microscopy staff provides strong support for the users with 2D and 3D image analysis. The Core’s Atomic Force microscope is heavily used to study molecular interactions with nanometer resolution, to evaluate the effect of inhibitors on pathologic processes, and to assess stiffness of tissues from normal and cancer-bearing animals as well as tissues from cancer patients.