Olympus FLUOVIEW FV300 - Page 44

Getting Started FLUOVIEW/Basic Operations
IV. OPERATION INSTRUCTIONS
IV . 1-6
Page
(5) Kr/Ar laser line filter switching (Available only with the Kr/Ar laser combination)
The 488 nm and 568 nm excitation wavelengths of the Kr/Ar laser can be selected
according to applications. This switch has 5 positions which can be selected with a
rotary knob.
1) 488: Excites the specimen at 488 nm. Use this position when observing
a single-labeled specimen such as FITC.
2) 568: Excites the specimen at 568 nm. Use this position when observing
a single-labeled specimen such as TRITC and PI.
3)568
+
488
AT6:
Excites the specimen at 488 nm and 568 nm, but the excitation
output at 488 nm is attenuated to 6% of the normal level. Use this
position to prevent interference of FITC emission on PI emission
when observing a double-labeled specimen such as FITC+PI.
4)568
+
488
AT25:
Excites the specimen at 488 nm and 568 nm, but the excitation
output at 488 nm is attenuated to 25% of the normal level. Use this
position to prevent interference of FITC emission on PI emission
when observing a double-labeled specimen such as FITC+PI.
5)568
+
488:
Excites the specimen at 488 nm and 568 nm. Use this position
when observing a double-labeled specimen such as FITC+PI.
The FITC emission tails longer than 570 nm. As a result, the FITC
emission tends to be detected by CH2 and interferes with the PI
emission (see diagram below). This problem may be solved ideally by
cutting off the FITC omission using OFFSET in the control panel
([Acquire] panel), but if this is not impossible, it is necessary to
attenuate the intensity of the 488 nm wavelength exciting the FITC
emission in order to balance the excitation emissions.
Otherwise, you can also use sequential scanning to obtain the same
result.
NOTE