Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
ZEISS Left Tool Area and Hardware Control Tools LSM 880
258 000000-2071-464 10/2014 V_01
The data of the
Count rate scan
window can either be copied to the clipboard or written to a text file. A
click in the Count rate scan window with the right mouse button will open the context menu for line or
two dimensional scans.
•
Select the line Copy data to clipboard with a click of the mouse if you want to insert the data into
other WINDOWS programs directly via the clipboard. The Paste function enables you to insert these
data directly into the required program.
•
Select Write text to file to save the data in an external ASCII file (.txt).
You can zoom in by spanning with the left mouse button held down a region of interest (not available
for a frame plot). You can rest the zoom by selecting the line plot by defining the zoom area with.
Z Scan for finding a labeled
cell membrane
•
Press the xyz-Scan FCS
action button to perform a z
scan. The z scan is performed
and the measured intensity is
displayed in dependence of
the z-position (Fig. 346). The
red line in the diagram shows
the actual z-position. Peaks
(normally two from the two
lower and upper membranes)
show levels of high signal
intensity and may correlate to
labeled membranes. Note,
that if the glass surface
happens to be in the z scan
range it will also give a peak.
To determine its position, just
perform a z scan at a position
where is no cell. If the glass
reflection contribute so much
that the peaks of the membranes are hidden due to scaling, you have to shorten the lower range end.
Place the red line at the peak that corresponds to the membrane to select this z-position. Note that z
scan is performed relative to the current position, which will be assigned to 0.
•
If no clear signal can be detected, or the peak of interest lies too close to the border, close the scan
window and perform a new z scan. Choose a different z range by modifying start and end positions to
values that lie closer to the position at which the signal should be expected.
•
If the z-position has been selected, close the
Z Scan
window by clicking on the
Close
button.
•
Press the Snap or Start Experiment action buttons to perform an FCS measurement. The FCS
measurement is performed at the same X/Y position as the z scan.
The z scan is especially appropriate to position the confocal volume on a cell membrane. Due to the
shape of the confocal volume, the membrane should not be approached from the side but instead rather
from the top of the cell. Please note that it is better to use the upper membrane for measurement, since
the lower membrane might be too close to the glass bottom surface resulting in disturbing reflections.
Besides using the
Z Scan
you can also position the membrane manually, albeit with less precision.
Fig. 346 Z Scan image