CONFOCOR
3
Carl Zeiss
System Configuration
ConfoCor 3
30 M60-1-0025
e
02/2010
Procedure for defining a new beam path configuration
•
First tick the laser or lasers (in case of cross -correlation) you need for excitation of your sample.
•
Then choose a major dichroic beam splitter which should correspond to the chosen laser(s).
•
Select an infrared block filter in case you need one with NLO applications.
Please note that you can use IR lasers for FCS, but there are no specifications for the performance
of the instrument for this excitation.
•
Select the secondary beam splitter.
−
For cross correlation, choose an appropriate filter like the NFT 488/633 beam splitter.
−
For autocorrelation measurements in detection channel 1 you select
Plate
.
−
All autocorrelation measurements in detection channel select
Mirror
.
•
Click the emission filters you need.
•
Activate the channels you want to use. It is recommended to give the channel a name which is easy to
remember.
•
Set the laser power.
•
Perform a Pinhole alignment.
•
Store beam path or method, which uses the beam path.
9.6.1.1
Pinhole
The
Pinhole
panel allows you to set the pinhole diameter and to automatically align the pinhole position
in x and y.
•
Click on the
Pinhole
tab to open the
Pinhole configuration
panel (see Fig. 35). Click on any other
register to leave the panel.
Setting the Pinhole diameter
In the
Pinhole
configuration
panel you can set the
pinhole diameter of the LSM using the Diameter
slider, scroll arrows or the input box (Fig. 35). The
display of the input box gives the diameter in µm.
The Pinhole diameter in Airy units is automatically
calculated and displayed beside the input box.
An optional iris in the excitation beam allows you
to enlarge the size of the confocal volume by
decreasing the pupil filling. If present a
Pupil
filling
slider and input box will be available in the
Pinhole menu (Fig. 35). You can enter a value by
typing, using the arrows or the slider. The Airy
units displayed right of the Diameter slider and
input box will be updated automatically.
Note if the system is switched to LSM mode, the
Fig. 35
Pinhole configuration panel
