Olympus IX2 RFAEVA2 - Manual (Page 16)

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Fig. 4
3-3 Observation Procedure
Fig. 3
1 Adjusting the Focus (Fig. 3)
1. Remove the specimen replacement lid @ from the stage.
2. Apply immersion oil on the objective top lens and place the specimen on
the stage.
3. Place the specimen replacement lid @ on the specimen box ²; the laser
beam will be output. When the image is displayed on the monitor for the TV
camera, bring the displayed specimen image into focus.
2 Adjusting the Laser Beam Incidence Angle (Fig. 4)
}Perform the laser beam incidence angle adjustment by referring to the
“Explanation Diagrams” on the next page.
}When the laser beam incidence angle adjustment knob 1 of the fiber
light illuminator is set to the fully clockwise position, the angle of the laser
beam output from the objective is smaller than the critical angle and total
internal reflection is not obtained (see Explanation 1).
# Do not turn the adjustment knob further clockwise from the above
condition by force.
1. While observing the monitored image, turn the adjustment knob 1 slowly
counterclockwise until total internal reflection fluorescence is obtained
(Explanation 3).
When the laser incidence angle approaches the critical angle (Ex-
planation 2), the bright part of the fluorescence image flows in the
vertical or horizontal direction of the field of view. When the angle
is increased further, the background fluorescence darkens sud-
denly and the total internal reflection fluorescence is obtained (Ex-
planation 3).
2. Turning the adjustment knob 1 further counterclockwise decreases the
range where total internal reflection fluorescence is observed (depth from
the cover glass surface). Turning the adjustment knob clockwise increase
the above range.
}If the adjustment knob is turned too much counterclockwise, nothing will
be visible eventually (Explanation 4).
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