OPERATION
Laser TIRF 3
Checking and Adjusting the Laser Beam Divergence and the TIRF Angle
Carl Zeiss
02/2014
423683-7144-001
77
4.5.2
Adjusting the Laser Beam
Divergence Using the Adjustment
Tool
1.
Focus on the glass/medium interface of the
specimen (adherent beads) in epi-fluorescence.
2.
Change to laser illumination and adjust 0°
angle of laser illumination in epi-fluorescence.
3.
After having focused the specimen by means
of the TIRF objective, change to the adjustment
tool in order to adjust the angle.
4.
Swivel the reflector module with the neutral-
density filter into the beam path.
5.
Observe the live image of the camera.
6.
Now, a white spot can be seen in the center of
the live image. This is the reflected laser beam.
7.
Adjust the camera so that the resulting images
are not overexposed.
8.
Change the TIRF angle so that the laser point
appears in the center of the live image.
9.
Now swivel the divergence adjustment tool
into the beam path.
−
You see now an enlarged image of the
laser beam profile.
−
Check the beam focus using the divergence adjusting screw of the Slider Laser TIRF.
−
A bright spot surrounded by two rings of lower intensity appearing in the center indicates
optimum adjustment (Fig. 4-6 "In focus").
−
Check the focus adjustments for the different laser lines of the existing laser modules.
−
If necessary, the beam focus for the laser lines used may be optimized this way.
10.
Finish the adjustment process by swiveling the TIRF objective into the beam path and selecting an
appropriate reflector module inserted in the reflector turret, or adapting the laser intensity again to
the previous value.
In focus
Out of focus
Far out of focus
Out of focus
Far out of focus
Huge diameter
Weak power
Fig. 4-6
Adjusting laser divergence
Summary of Contents for TIRF 3
Page 1: ...Laser TIRF 3 Imaging System Operating Manual...
Page 110: ......
