Olympus IX2-ARCEVA - Manual (Page 17)

IX2-ARCEVA
1 4
4-2 Total Reflection Fluorescence Observation
· The only objective that can be used in total reflection fluorescence observation is the PlanApoN60XO TIRFM or PlanApo60XO TIRFM .
· Since the illumination intensity is lower than with the laser light source, this system is not applicable to molecular
fluorescence observation. Its applications are limited to high-contrast observations of tissues near cell membranes and
physiochemical phenomena.
1. Set the slit to the OUT position and the wedge prism to the pulled out vacant position.
2. Using an objective other than the PlanApoN60XO
TIRFM or PlanApo60XO TIRFM , set up the microscope condition for general
fluorescence observation.
3. Push in the field iris diaphragm lever to the open condition and engage PlanApoN60XO
TIRFM
or PlanApo60XO
TIRFM
objec-
tive in the light path.
4. Focus on the lowest position of the specimen (position closest to the cover glass).
5. Set the wedge prism in the intermediate click position and push in the slit to the IN position.
Now the microscope is ready for total reflection fluorescence observation.
The effusion depth can be fine adjusted by slightly rotating the lower slit centering screw as described below.
· Clockwise:
This moves the slit inward, increasing the effusion depth.
· Counterclockwise: This moves the slit outward, decreasing the effusion depth.