Zeiss LSM 880, Operating Manual

Page: 192 / 690

CHAPTER 1 - SYSTEM OPERATION
ZEISS Left Tool Area and Hardware Control Tools LSM 880
186 000000-2071-464 10/2014 V_01
5.3.2.5 Pinhole Adjustment
The Adjust Pinhole panel allows you to set the pinhole diameter and to automatically align the pinhole
position in x and y.
•
Click on the Adjust Pinhole tab to open the Pinhole configuration panel (Fig. 269). Click on Close
to leave the panel.
Setting the Pinhole diameter
Activate the channel you want to use and
that is appropriate for your settings by
pressing the respective
Ch tab. In one
channel settings only the active used channel
will be available. In two or more channel
settings both channels will be available. You
can adjust the pinhole to either channel. The
values for the pinhole alignment done with
the last channel will be stored and used with
a method.
You can set the pinhole diameter of the LSM using
the Diameter slider, scroll arrows or the input box
(Fig. 269). The display of the input box gives the
diameter in µm. The Pinhole diameter in Airy units
is calculated automatically and displayed beside the
input box. The
1 AU
button allows you to set the
pinhole size to 1 Airy unit by just clicking on it. Set
the desired pinhole diameter using the slider, the
scroll arrows or type into the input box.
This will alter the pinhole diameter automatically also for the method in the
Pinhole
panel. If you
need another pinhole size for doing the pinhole alignment be sure to reset it before saving the
method.
•
It is recommended to set the pinhole diameter to a size corresponding to the used excitation
wavelength that corresponds to 1 AIRY unit:
−
458 nm: 33 µm
−
488 nm: 34 µm
−
514 nm: 39 µm
−
561 nm: 40 µm
−
633 nm: 45 µm
Pinhole sizes are given in µm and AIRY units.
Please note that there is only one pinhole for all detector channels of the various modules. NDD
does not have a pinhole. You can adjust the pinhole to either channel. The values for the pinhole
alignment done with the last channel will be stored along with the beam path or a method and
used. A method is always saved together with the system configuration, acquisition and processing
parameters. A beam path is saved only with the pinhole settings.
You can adjust the beam path to the pinhole of the LSM manually or automatically. Manual adjustment is
not recommended, since the relevant procedure is very complex.
Fig. 269 Adjust Pinhole panel