In Vivo Fluorescence Microscopy of GFP-Labeled Cells

For the in vivo fluorescence microscopy of cells expressing pJMCS-DM23 (GFP-2XPH-PLCδ1) [36 (link)], exponentially growing SR-G cultures treated or not with neomycin 5 mg/mL were collected by centrifugation at 5000 rpm for 1 min and directly observed with an Eclipse TE2000U microscope (Nikon, Tokyo, Japan) using the appropriate sets of filters. For pRS410-GFP-LactC2 observation, the same procedure was followed but cells were cultured in SD. Digital images were acquired with an Orca C4742-95-12ER charge-coupled device camera (Hamamatsu, Hamamatsu city, Japan) and processed with HCImage software (Hamamatsu). Statistics on cell populations were performed by counting >100 cells for each experiment.