Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
LSM 880 Left Tool Area and Hardware Control Tools ZEISS
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(1) Manual Focusing on the Membrane
•
Acquire an image of the cell.
•
Place the focus above the cell and scan continuously.
•
Focus slowly down until you see a fluorescence signal.
•
Stop focusing.
If the signal is too low for imaging, the membrane can still be detected using the Z Scan feature.
(2) Focusing on the Membrane by a Z Scan
The Z Scan measures the count rate at previously defined x, y positions. It is performed without table
movement, which is at the fixed position of the laser beam.
•
Acquire an image of the cell, focus through manually and select an equatorial section (middle of cell).
Optional: Perform a
Z-Stack that includes the membranes of the cell with the LSM and select an
equatorial section.
•
Go to the
Measure
tool to
Acquisition
and the
Positions
submenu. Position the region of interest at
the site of the fixed laser beam in Current Position using the Crosshair or use the crossline in the
LSM image
to mark the position. In the latter case,
Scanner
must be active, since the
XY-Stage
will
approach the marked position only during a measurement!
•
Go to the
X-Y-Z Scan
tool and activate
Z
. The
Z Scan
display shows the current z-position of the laser
beam.
•
Select the
Start
and
End Position
and enter the
Step Distance
. If you have chosen an equatorial
section going 10 µm up and 10 mm down from the displayed current position are good values to start
with. Also a step distance of 0.5 µm should work well.
5.3.19 Snap
If the Snap button is pressed, a measurement is triggered according to the System Configuration ,
Acquisition
and
Correlator settings
.
However, in regard to positions Snap triggers the measurement only at the highlighted or current
position.
5.3.20 Start Experiment
If the Start Experiment button is pressed, a measurement is triggered according to the System
Configuration
,
Acquisition
and
Correlator settings
.
All selected positions will be approached sequentially.