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Fluorescence Microscopy
“fluorescence imaging has become the
mainstay of microscopy” -- Nat. Methods, 2005
Objectives:
1. Understand the basics of fluorescence
2. Understand how fluorophores are used in
microscopy
3. Understand how a microscope is set up to do
fluorescence
4. Understand basics of immunofluorescence
Have you used a fluorescence microscope
before?
A. Yes
B. No
Fluorescence microscopy uses substances
called “fluorophores” or “fluorochromes”
as stains.
These substances have the property that
they “fluoresce”, i.e. they absorb some
wavelength of light, become excited, and
decay to the ground state by emitting a
slightly longer wavelength of light.
R O Y G B I V
700 650 600 550 500 450 400
wavelengths in nm
E = h
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