Quick Microscope Reference:
Note: In order to properly set up phase-contrast microscopy, or fluorescent
microscopy, you will need to adjust the settings to optimize “bright field” prior to
any other adjustments. I will refer to it as Köhler illumination rather than bright
field, as bright field is only one application that KI can be applied.
Köhler illumination sets up the microscope for the optimal optical path taking both
the filament alignment as well as the condenser alignment into consideration. Both
of these must be properly aligned for optimal photomicroscopy.
For a quick tutorial, you can use the Nikon U site:
http://www.microscopyu.com/tutorials/java/kohler/
Phase Contrast depends on the diffraction of light through the tissue. This light is
segregated and reflected back producing the high contrast image seen. Proper
alignment of the Objective Rear Focal Plane is crucial to phase imaging, as well as
the proper Köhler illumination setup.
Köhler Illumination
1. Release the condenser by turning the condenser refocusing clamp
2. Set the eyepiece turret to <O>
3.