Chapter 2 Microscopy Techniques
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2.2 Phase Contrast Microscopy
Phase contrast microscopy is a method of using diascopic illumination to observe unstained, colorless and
transparent specimens such as live cells.
Phase contrast microscopy uses the interference between direct light and light diffracted while passing
through a colorless and transparent specimen to visualize light phase shifts as brightness variations.
Insert the phase contrast slider with the annular diaphragm into the slot on the side of the condenser, and
bring the phase contrast objective with the same PH code into the optical path. The microscope is now ready
for phase contrast microscopy.
For details on the configuration and the assembly of the microscope for phase contrast microscopy, see “4.2.2
Configuration for Phase Contrast Microscopy.”
1
Prepare the microscope.
Check that the following optical parts are usable.
(1) Phase contrast slider
1lider for 10x-40x:
Supports 10x and 40x Ph
objectives. When switching to phase contrast
microscopy, remove the dummy slider from the slot
on the side of the condenser and insert this slider.
Slider for 100x:
Supports 100x Ph objectives. An
empty position is available, so bring the empty
position into the optical path in advance. When
switching to phase contrast microscopy, operate the
slider to bring the phase contrast position into the
optical path.
(2) Phase contrast objective (Ph objective)
Check that the Ph objective is attached to the
nosepiece.
Combination of optical parts
Confirm in advance that the slider is paired with a
supported objective with a supported PH code.
(3) GIF filter (green interference filter)
Attach the GIF filter to the field lens when performing
phase contrast microscopy.
(4) Centering telescope
Use the centering telescope when adjusting the
annular diaphragm position.
(3) GIF filter
(4) Centering telescope
2
Focus on the target in bright field microscopy.
Follow steps 1 to 12 of the procedure in “2.1
Bright-field Microscopy”, and focus on the target
using the 10x objective with the bright-field
microscopy method.
(1) Phase contrast slider
(for 10x-40x, for 100x)
(2) Phase contrast
objective (Ph objective)
