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FLIM Data Acquisition
3. Deactivate all PMTs.
4. Activate the APDs by clicking the corresponding buttons ( Figure 115, item 2 ).
5. Select low laser intensities.
6. You may also have to adapt the color look-up table (LUT). In the dialog of the respective
detector, click the color scale to call up the LUT Selection dialog. There, you can select
color look-up tables (LUT) for displaying the images to be acquired and create user-
defined color look-up tables.
You can also configure the intensity setting via
Gain
and the control panel. This function
has no effect on the detector; it modifies the color look-up table, which renders low-
intensity structures more visible. It may also be helpful to accumulate over several
images. The
Offset setting on the control panel has no effect on the image.
7. Optimize the image quality. The functions for this are the same as outside the wizard.
8. Acquire an image by clicking the Capture Image image acquisition button or acquire a
stack by clicking the
Start
image acquisition button. The start and finish of the stack are
defined interactively. The stack image is automatically added to your LAS AF experiment.
16.1.1.3 External FLIM Photomultiplier
1. For light to fall on the detector, select the
---
option for
X1 Port
.
2. You can enable the PMT Extern option in Beam Path Settings . It is required for
automated functions in the FLIM Wizard (
Setup FLIM
and
Measurements
steps).
Activating it causes empty images to be created in LAS AF.
16.1.2 Selecting Laser Lines as an Excitation Source for Image Acquisition
All available lasers can be used for the image acquisition. You will find a description of the
mode of operation in
Chapter 15 .
16.1.2.1 Using Continuous Wave Lasers
1. For capturing the emitted fluorescence, set the acousto-optical beam splitter to
fluorescence ( Figure 116 ).
Dependency of the spectral detection range of the APDs
The spectral detection range of the APDs is determined by the SMD filter
cube used, not by the slider settings for the photomultipliers.
Images of the external FLIM PMT only in SymPhoTime
The signal of the external FLIM PMT is not displayed in LAS AF. You can
view images created by this detector only in SymPhoTime. The
corresponding detector displayed in LAS AF is a simulator required for
the function of the wizard.