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10.4.
FRAP
1. Click on
Edit Bleach
toolbar button to open up
Bleach Control
dialog.
2. Set Bleach parameters in
Bleach Control
dialog
Check
Bleach after Number
of Scans
Set “
Scan Number
” to for pre-bleach imaging
Set “
Iterations
” value – this may require empirical
determination. It needs to be enough to allow complete
bleaching but few enough to allow quick return to imaging.
Try 10 as a first attempt.
In “
Excitation of Bleach Track”
set bleaching laser to
100%
3. Click
Define
Region
button to select
bleach area
4. Define area to bleach then select it by
ticking the
checkbox
along side the ROI.
Close dialog.
5. Open
Time Series Control
dialog by clicking the
TimeSeries
button in the main toolbar.
5. Set FRAP time-course in
Time Series Control
Dialog
Start Series
=
Manual
.
Stop Series
= select
Time
then enter the duration of the
experiment.
Enter
Cycle Delay
(time between frames).
7. Click “
StartB
”
to start experiment.
