Last Updated on Sep 2021
4.6 Setting Laser & Detector configuration
•
Based on the no. of dyes, you have selected, it will give you
the total no. of seq. (E.g., if you have selected 3 dyes in
sequential mode, you will have 3 seqs corresponding to each dye.)
•
Select each seq, and increase the laser power to 1% initially. Make sure the laser line is
activated by checking both the red button and the laser line shown at 2 and 3.
•
Make sure that the HyD/PMT is activated. (Turn on the slider)
•
Then, Select the fluorophore you are using. It will give you the emission spectrum of the
dye you are using.
•
Choose an appropriate emission band to detect your fluorophore. (Important: Make sure
that the emission band is at least 8-10nm away from your excitation line.)
•
Choose an appropriate detector pseudocolour (also known as a Look Up Table, or LUT).
•
For PMT, set the Gain to approx. 700 - 800 and the Offset to 0.
•
When using the HyD (2× more senstitive than PMT), default gain is set at 100
.
(Important:
Do not overload HyD, max is 100.)
•
Gating: Removes the first few nanoseconds of the signal as a result of autofluorescence
from the sample or mounting medium.
o
Only usable on pulsed lasers such as the WLL. Constant mode lasers like the Argon lasers
will not work with the Gating function.
o
Choose appropriate reference laser line for gating.
o
NOTE:
start with low laser power to avoid damaging the HyD detectors. HyD detectors
will shut down if an overload condition is detected.
1
2
3
Gating
Ref.line
