Figure 2. HEp-2 cells were stained with ANA antibody (BIO-RAD) and with the indicated fluorophores conjugated to comparable secondary antibodies. For fluorescence microscopy cells were mounted in ProLong Gold (Invitrogen) (upper images) or PBS-90% glycerol containing n-propyl gallate (Fluka) as an anti-fading agent (lower images). Images were taken at 100X with a Nikon D-70 DSLR camera connected to a Nikon LABOPHOT epi-fluorescence microscope. Fields with the brightest cells for each fluorophore were selected for imaging after 1 second exposure to light.