Leica TCS SP8 SMD, User Manual

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F(L)CS Data Acquisition
18.2 Setup Imaging Step – Image Acquisition
1. Start the FCS Wizard in LAS AF. To do so, open the operating mode selector and select
FCS .
2. For applications in cells or tissues, in the
Setup Imaging
step you first create an xy scan
of your specimen in the desired z plane, or you acquire an xyz stack (with z-Galvo).
3. If you would like to display a 'z-section' of your specimen, you can also acquire an xz
image or an xzy stack. This might be especially of interest when working on membranes.
Other imaging modes (like time series or lambda stack) are not supported. The image
acquisition procedure is the same as the general procedure without using the FCS Wizard.
18.2.1 Selecting Detectors for the Image Acquisition
You can use photomultipliers as well as APDs for the image acquisition. Photomultipliers are
distinguished by having wide dynamic range; APDs are distinguished by higher detection
quantum efficiency. PE APDs are distinguished by very good red sensitivity.
18.2.1.1 Photomultiplier (PMT) / Internal Hybrid Detector (HyD)
1. Check that the external port X1 is switched to the Mirror position ( Figure 166, item 3 ).
2. In the beam path schema, activate the desired PMTs/HyD by clicking the appropriate
check box.
3. Select the desired spectral range for detection.
4. Select low laser intensities.
5. You may also have to adapt the color look-up table (LUT). 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.
6. Optimize the image quality. The functions for this are the same as outside the wizard.
7. Acquire an image by clicking the
Capture
image acquisition button or acquire an xyz or
xzy stack by clicking the Start image acquisition button. The start and finish of the stack
are defined interactively, in the usual way. The stack image is automatically added to
your LAS AF experiment.
18.2.1.2 PE / MPD APDs
1. First, switch on the APDs on the APD or DSN control unit electrically.
2. Set the external port X1 (
X1 Port
) to the position
---
(
Figure 168, item 1
).
3. Deactivate all photomultipliers in Beam Path Settings .
4. Activate the APDs by clicking the corresponding check boxes (
Figure 168, item 2
).
5. Select low laser intensities.
6. You may also have to adapt the color look-up table (LUT). 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