LSM 880
Left Tool Area and Hardware Control Tools
ZEISS
10/2014 V_01
000000-2071-464
171
5.2.23.2
Defining the Scan Parameters
•
Open the
Acquisition Mode
tool (Fig. 252).
•
Select
Frame
mode and a
Frame Size
of 512 x
512 to start with.
•
The
Optimal
button automatically adjusts the
frame size to match the magnification and
resolution of the objective. Thus, images are
acquired without collecting redundant image
information.
•
The Scan Speed is set to 9 per default. A lower
speed produces better images as the light
collection per pixel is higher. Use a speed from
7 to 9 to begin with.
•
Start with the following settings on the
Averaging
panel:
Bit depth:
8 bits
Number:
1 (no averaging)
Additional parameters like scan direction are displayed in
Show all
mode. Scan direction is per
default set to unidirectional.
The default settings for Acquisition Mode are suitable for good image quality and do not need
to be changed for initial scanning.
5.2.23.3
Defining the Detection Parameters
•
Activate the
Channels
tool (Fig. 253).
•
Set the
Pinhole
to one Airy Unit clicking 1 AU.
−
The pinhole diameter should be so small that
there is still enough variation for the setting
of the Gain (Master) and that sufficient
image information is still available. 1 Airy is a
good value to enable a confocal fluorescence
XY-image to be obtained.
−
A small pinhole diameter will increase the
depth of focus, but reduce the light intensity
received by the PMT photomultiplier (for
reflection mode confocal images start with a
pinhole value of 0.5 Airy Units).
•
Use the
Set Exposure
button.
−
The system determines the settings for the
Gain (Master) and digital offset to match the
scan speed, pinhole size and laser power.
−
Finally, the system builds up the image.
Fig. 252
Acquisition Mode tool
Fig. 253
Channels tool
Summary of Contents for LSM 880
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