Imaging Platelets and Megakaryocytes by High-Resolution Laser Fluorescence Microscopy

Microscopy is central to studies of platelets and their precursor megakaryocytes. Here we describe methods to rapidly obtain high resolution images of fixed platelets, megakaryocytes and megakaryocytic cells via immunofluorescence microscopy. Protocols covered include: (1) isolation and preparation of cells suitable for fluorescence staining; (2) staining with antibodies and other molecules; (3) imaging via spinning-disc confocal and structured illumination laser fluorescence microscopy; (4) processing and presentation of images. Also included is a list of primary antibodies we have validated for use in staining specific proteins and subcellular structures in platelets and megakaryocytes.