Chapter 2 Microscopy Operations
76
Phase Contrast
Microscopy
Preparation
Focus and Optical
System Adjustment
Microscopy
Phase Contrast
Microscopy Procedure
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14
Center the PH module (Ph annular diaphragm).
To optimize the phase contrast effect, it is important to
properly overlap the phase plate of the objective with the
PH module (Ph annular diaphragm) image in the
condenser.
(1) Make sure that the 10x objective (Ph1) and position
[Ph1] of the condenser turret are in the optical path.
(2) Rotate the stage knob to move the specimen and bring
a portion where there is no sample under the cover
glass into the optical path.
(3) Remove one eyepiece from the tube, and attach the
centering telescope in place.
(4) Hold the flange of the centering telescope and rotate
the eyepiece to focus on the phase plate of the
objective.
(5) If the phase plate of the objective is not aligned with the
PH module (Ph annular diaphragm) of the condenser,
align them by raising and pushing the two PH module
centering knobs on both sides of the back of the
condenser and turning the centering screws. When
done, restore the centering knobs to the original
position.
(6) Place the eyepiece back into the tube.
Centering of the Ph annular diaphragm
and the phase plate
The phase image will slightly differ depending on how
the Ph annular diaphragm for the PH module
overlaps the phase plate for the objective. For a
precise observation or capturing of still images,
check whether the annular diaphragm and the phase
plate are concentric at each magnification. (See “18.1
“Tips for Phase Contrast Microscopy” - “Centering of
the objective (phase plate) and the PH module (Ph
annular diaphragm)”.)
Centering the PH module
Bad Good
Centering telescope
Eyepiece
Flange
PH module
centering knob
(on both sides)
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