SIMULTANEOUS FLUORESCENCE OBSERVATION
METHODS
}By properly combining equipment, this system can be used in transmitted light brightfield observation, transmitted light phase contrast observation and transmitted light DIC (Differential Interference Contrast) 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, making it easy to tell which portion of the specimen is fluorescing.
}The IX81 and IX71 have a lamp ON-OFF button. Use this button to switch combined observations to the fluorescence observation only.
1
Simultaneous Reflected Fluorescence and Phase Contrast Observations
}The phase contrast observation requires the IX2-LWUCDA2, IX2-LWUCD, IX-ULWCD, IX2-MLWCD or U-UCD8 condenser and a Ph objective.
1. Engage a dummy mirror unit (an empty position) 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 in the light path and release the shutter.
5. Adjust the transmitted light for the best balance of fluorescence and phase contrast brightness.
}Use ND filters or green filter in combination to adjust the transmitted light intensity. This can also be adjusted with the light intensity control of the microscope frame.
Remember that the service life of a halogen bulb tends to decrease when the bulb is kept on continuously at a low voltage when it is to be turned off, so be careful.
2
Simultaneous Reflected Fluorescence and DIC (Transmitted Light) Observations
}The DIC observation requires the following modules.
· Condenser (IX2-LWUCDA2, IX2-LWUCD, IX2-DICD, IX2-MLWCD or U-UCD8)
· DIC slider (U-DICT, U-DICTS, U-DICTHC or U-DICTHR)
· Analyzer (U-ANT or IX2-AN)
# In order for reflected fluorescence to be effective in the simultaneous observation, insert the IX2-AN analyzer into the analyzer insertion slot below the dichroic mirror.
Do not insert the U-ANT analyzer in the DIC slider, for this will dim the fluorescence observation imaged and cause the analyzer to be burnt.
1. Engage the dummy mirror unit (on an empty position) in the light path.
2. Adjust the polarizer on the DIC condenser to the “Crossed Nicol” (complete extinction) position.
3. Insert the DIC slider into the position provided on the revolving nosepiece.
4. Rotate the turret on the condenser to select the Nomarski prism matching the objective 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 and aperture iris diaphragm.
8. Turn the prism movement knob on the DIC slider to adjust contrast of the DIC image.
9. Engage the mirror unit corresponding to the desired excitation and release the shutter.
10.
Adjust the transmitted light for optimum fluorescence and DIC image brightness.
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