Page 220 - Fundamentals of Light Microscopy and Electronic Imaging

EXAMINING FLUORESCENT MOLECULES IN LIVING CELLS 203

spectra of fluorescein and rhodamine (Figs. 11-6, 11-8 and Color Plate
11-1).
(3) Is rhodamine fluorescence totally excluded from the fluorescein filter set
with the narrow band-pass filter? Why? Since the long-pass fluorescein
filter set shows transmission of red wavelengths, it should be obvious that
rhodamine fluorescence will also be transmitted by this filter set design,
making it unsuitable for examination of double stained specimens.
(4) What is the purpose of equipping a microscope with a filter set containing
a long-pass emission filter?
(5) Name two steps you could take to reduce the amount of rhodamine bleed-
through in a fluorescein filter set.