Olympus U-RFL-T Manuel d'instructions - Page 21
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SIMULTANEOUS FLUORESCENCE OBSERVATIONS
} By properly combining equipment, this system can be used in transmitted light brightfield observation, transmitted phase
contrast observation and transmitted light DIC observation in addition to the reflected fluorescence observation. With
specimens that fade rapidly, fading can be minimized by initially using transmitted light phase contrast or transmitted
light DIC observation for positioning. Reflected fluorescence observation can also be executed simultaneously with phase
contrast or DIC observation.
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Simultaneous Reflected Fluorescence and Phase Contrast Observations
} For details on using phase contrast observation, refer to the instruction manual provided with the phase contrast condenser
or universal condenser.
The phase contrast observation requires a phase contrast condenser (U-PCD2) or a universal condenser (U-UCD8) and
a Ph objective.
1. Engage a dummy mirror unit (or an empty position on the turret) in the light path.
2. Rotate the phase contrast turret to show the same number as the Ph number shown on the objective.
3. Adjust the optical axis between the ring slit and phase plate by centering them.
4. Engage the mirror unit corresponding to the desired excitation into the light path and open the shutter.
5. Adjust the transmitted light for optimum balance of fluorescence and phase contrast brightness, and you are ready for
observation.
} To adjust the brightness of transmitted light, Use ND filters or adjust the transmitted light intensity.
Simultaneous Reflected Fluorescence and Transmitted Light Differential
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Interference Contrast (DIC) Observations
} For details on the transmitted light DIC observation, refer to the instruction manual provided with the U-UCD8 transmitted
light universal condenser.
The transmitted light DIC observation requires the following accessories; @ universal condenser (U-UCD8); 2 transmitted
light DIC slider (U-DICT, U-DICTS, U-DICTHR or U-DICTHC); 3 analyzer (U-AN-2); | revolving nosepiece for DIC (U-D6RE,
U-D7RE or U-D7RES).
} In order for reflected fluorescence to be effective in the simultaneous observation, insert the U-AN-2 analyzer into the
analyzer insertion slot above the dichroic mirror on the illuminator.
1. Engage the dummy mirror unit in the light path.
2. Adjust the polarizer on the universal condenser to the "crossed Nicol" (complete extinction) status.
3. Insert the transmitted light DIC slider into the position provided on the nosepiece.
4. Rotate the turret on the universal condenser to select the Nomarski prism matching the objective and DIC slider to be
used for observation.
5. Engage the objective to be used in the light path.
6. Place the specimen on the stage and focus on the specimen.
7. Adjust the field iris diaphragm of the transmitted light illumination unit (built into the microscope base) and the aperture
iris diaphragm of the universal condenser.
8. Turn the prism movement knob on the transmitted light DIC slider to adjust contrast of the DIC image.
9. Engage the mirror unit corresponding to the desired excitation in the light path and open the shutter.
10. Adjust the transmitted light intensity for optimum fluorescence and DIC image brightness.
Note
} if you are frequency switching between reflected fluorescence observation and transmitted light Nomarski DIC observation
but you do not need to use both simultaneously, then it will be more convenient for you to use the DIC mirror unit (U-FDICT)
instead of an analyzer (U-AN-2). This facilitates the switching operation because the analyzer simultaneously enters the
light path when the fluorescence mirror unit is switched to the DIC mirror unit.
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