OPERATION
Laser TIRF 3
Adjusting the TIRF Effect on the TIRF Slider man.
Carl Zeiss
02/2014
423683-7144-001
79
4.6
Adjusting the TIRF Effect on the TIRF Slider man.
•
Place a specimen in the object mounting frame.
•
Move the required TIRF objective into the optical path.
•
Apply immersion oil to the front lens system of the objective
TIRF microscopy must be performed in an aqueous medium, with a refractive index in the range
from n = 1.33 to 1.38!
The TIRF effect does not occur with specimens embedded, for instance, in Canada balsam
(n = 1.51).
For pre-adjusting the Laser TIRF imaging system it is advisable to carry out an experiment with
fluorescing latex beads which enables you to observe very clearly the occurrence of total
reflection. This is particularly beneficial for multicolor TIRF, too. Therefore, move the beads or
the object to be examined into observation position. Focus on the surface of the cover slip
either in transmitted-light or epi-fluorescence illumination. Fluorescence allows you to identify
this position due to the immobilized beads. The evanescent field will be created in this focal
plane when adjusting the TIRF angle.
Fig. 4-8
Adjusting the TIRF effect
•
Turn on the reflected light source as per the operating instructions. Wait for approx. 10 minutes until
even illumination is achieved.
•
Once the Slider Laser TIRF is ready for operation, the
Active
4
) and
Safe
3
) LEDs
light up.
•
Activate epi-fluorescence illumination, if desired, by pressing the
RL
button on the right-hand side of
the microscope stand.
•
Focus the specimen (beads) on the cover slip surface using the rotary button (Fig. 4-8/
1
).
Summary of Contents for TIRF 3
Page 1: ...Laser TIRF 3 Imaging System Operating Manual...
Page 110: ......
