Optimized Transillumination
in the Microscope
"Correct illumination of the microspecimen is the single most important
aspect of critical microscopy and photomicroscopy. Fully 80 percent of all
photomicrographs submitted for contests and exhibitions are rejected because of
improper illumination of the microspecimen. Another 10 percent of the
photomicrographs reflect improperly adjusted aperture diaphragms".
[Photography through the Microscope, Eastman Kodak Company, 1988, Ninth Edition]
Dr. August Köhler published a paper in 1893 that described a method to
obtain the highest intensity of even illumination from a nonhomogeneous source.
Applying the "Köhler" method requires the control of 2 iris
diaphragms in the light path: the Field and the Aperture. The Field eliminates
all stray light by restricting illumination to the actual area under
observation. The Aperture controls the cone of light transilluminating the
sample so it fills the aperture of the microscope objective, thus optimizing
depth of focus and image contrast.
The relationship of diaphragm position and objective lens magnification can
be seen above. In an incident light system, the light is directed to the back of
the objective lenses. In the transmitted light system, the light is directed to
a substage condenser lens with a numerical aperture that approximates the
highest numerical aperture of the objective lenses. The substage condensor is
automatically lowered to expand the illumination field when low numerical
aperture objective lenses are in use. The entire assembly is motorized to
provide remote focus capability. The PC-Scope is calibrated to adjust the
diaphragms to their proper settings automatically as a function of objective
lens selection.
|