OPERATION
Laser TIRF 3
Controlling the Laser TIRF Imaging System Using ZEN 2012 (Blue Edition)
Carl Zeiss
02/2014
423683-7144-001
81
4.7
Controlling the Laser TIRF Imaging System Using ZEN 2012 (Blue Edition)
The basis of the following outline of how to integrate the TIRF imaging system laser into the ZEN 2012
software (blue edition) is the corresponding software manual or the online help function of the program.
This section focuses exclusively on providing a few useful pointers regarding calibration of the system.
Control of the TIRF imaging system laser is fully integrated into the software concept of ZEN 2012. Once
the system has been configured using the MTB 2011 configuration program, the laser module is available
as a light source.
Use of the Slider Laser TIRF mot. allows numerical data on the illumination angle used for each color
channel of a captured image and the resulting theoretical penetration depth to be saved.
It is necessary to calibrate the Laser TIRF Imaging System before actually using it. The purpose of
calibration is to render the illumination angle and the desired penetration depth reproducible. This
basically means that, after the calibration process the user should obtain a comparably bright image of
this sample for a given illumination angle, regardless of the side from which the sample is illuminated.
The calibration process compensates for any tolerances in the opto-mechanical structure of the system
and combines them into an offset which is taken into consideration by the software when controlling the
Slider Laser TIRF mot. Calibration is performed once the system has warmed up. The imaging software
includes a calibration wizard for this. Calibration can be carried out either using a test sample (fluorescing
latex beads) or on the specimen itself. If the actual specimen is used, the user should first ensure that
adherent fluorescence signals are available in the sample for focusing, otherwise inaccurate offset values
could be calculated.
Once these preparatory steps have been completed and an experiment set up, the user merely has to
select the desired laser line for the desired channel as a light source for the experiment.
Summary of Contents for TIRF 3
Page 1: ...Laser TIRF 3 Imaging System Operating Manual...
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