============================================ Live nuclear staining using Hoechst ============================================ Materials ---------- ============================================================================================== ========================= **Stock solutions** **Working solution** ============================================================================================== ========================= `Hoechst 33342 Solution `_ (20 mM) Dilute 1:10,000 in media ============================================================================================== ========================= Protocol ---------- .. time:: 30 min total 1. Remove media and add Hoechst working solution, diluted using the media of the cells in culture. A half-volume (e.g., 50 µL/well in a 96-well plate) can be used. 2. Incubate cells at 37°C for 20 min. 3. Aspirate the Hoechst solution and replace with appropriate culture media or PBS. 4. Image stained nuclei using the Keyence microscope or analyze cells by flow on the Attune.