Defining The Scan Parameters; Defining The Detection Parameters - Zeiss LSM 880 Operating Manual

LSM 880
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:
Number:
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.
−
10/2014 V_01
CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
8 bits
1 (no averaging)
000000-2071-464
Fig. 252 Acquisition Mode tool
Fig. 253 Channels tool
ZEISS
171