Tools In The Fret View Options Control Block For Acceptor Photobleaching - Zeiss LSM 880 Operating Manual

LSM 880 Center Screen Area / Image Containers - Display and Image Analysis
6.18.1 Tools in the FRET View Options
Control
Photobleaching
FRET tab (Fig. 695):
Method
As for
Photobleach is automatically selected in the drop
down list.
Within this tab the overlay regions present in the
image (defined as bleach, background or control
region of the bleach experiment) can be activated
(The drawing tools correspond to the drawing
tools described in section Graphics). Additional
regions can be defined for analysis as well.
The checkboxes Numbers and Measure refer to the overlay regions and annotate the number of the
region and its area to the overlay in the image.
Export allows the resulting FRET image to be saved as a separate image file.
The regions defined can be set as Object (Region of which FRET should be calculated) or Background
(from which the threshold for the image analysis is deduced from and automatically used for analysis and
displayed in the Threshold tab).
Regions and the assigned status as Object or Background can be selected or deselected for the individual
analysis using the corresponding check box under Enabled.
To start analyzing the Analyze button must be clicked once. If all further changes shall be effective
immediately the On the fly checkbox can be checked.
Parameter tab (Fig. 696):
Various parameters of the image series can be set
in this tab.
The image of the Donor and the Acceptor are
assigned to the appropriate channel using the
drop down menu next
Acceptor ch.:
The frame of the time series after the bleaching
event (Bleach after frame) is set using the slider
next to it or by editing the field next to the slider. If
only one bleach event was performed, this defines
the frame before and after the bleaching
respectively for analysis.
In case of a bleach experiment with repeated bleaching time points the checkbox Bleach Repeat must
be checked. This results in activation of the Donor post-bleach Frame slider and field. Here the frame
can be chosen to provide the data of the donor intensity after bleaching (D post) for analysis.
10/2014 V_01
CHAPTER 1 - SYSTEM OPERATION
Block for Acceptor
Acceptor
FRET Analysis,
Donor ch.:
to
000000-2071-464
Fig. 695 FRET View Options Control Block,
FRET tab
and
Fig. 696 FRET View Options Control Block,
Parameter tab
ZEISS
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