Use DAPI/HOECHST at the same concentration you use for cells/FS I use Hoechst H33258 Sigma 10mg/ml soln I dilute by adding 0.5 microL to 100 microL buffer then add 1/100 to Alexa secondaries ( final 1/20K diln factor) Some incubate in HOECHST or DAPI after secondary ab incubation, for IF Sure, if only "staining" nuclei, incubate in 1/20K Hoechst in same buffer This will vary for each lab Or, buy mounting medium that contains DAPI
Good luck Carl Carl Hobbs FIBMS Histology and Imaging Manager Wolfson CARD Guys Campus, London Bridge Kings College London London SE1 1UL 020 7848 6813 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet