OPERATION
Axioskop 40
Carl Zeiss
Illumination and contrast procedures
Axioskop 40 FL
3-34
B 40-810 e 12/01
3.5.7
Setting epi-fluorescence
(1)
General principle
The epi-fluorescence technique enables high contrast images of fluorescent substances to be displayed in
typical fluorescence colors. In the epi-fluorescence microscope, light generated by a high-performance
illuminator reaches the excitation filter (band pass) via a heat protection filter. The filtered, short-wave
excitation emission is reflected by a dichroic beam splitter and focused on the specimen via the objective.
The specimen absorbs the short-wave emission and then emits the long-wave fluorescence (Stoke’s law),
which is now gathered by the objective and transmitted by the dichroic beam splitter. Finally, the rays
pass a barrier filter (long pass/band pass), which only allows the long-wave emission from the specimen
to be transmitted.
Excitation and barrier filters, which are both positioned in the FL P&C reflector module together with the
corresponding dichroic beam splitter, must be perfectly matched.
(2)
Axioskop 40 FL configuration
−
Recommended objectives: Plan-Neofluar or Fluar (UV-excitation)
−
FL P&C reflector module and shutter plate in the reflector turret
−
HBO 103 or HBO 50 mercury vapor short-arc lamp for incident-light illumination
−
Fluorescence protection shield
☞
Before the epi-fluorescence technique is used, it is absolutely essential to adjust the mercury
vapor short-arc lamp. If required, re-adjustment must be performed depending on the
operation time.
(3)
Setting epi-fluorescence on the Axioskop 40 FL
The first epi-fluorescence setting is considerably simplified if the Plan-Neofluar objective 20x/0.50 and a
specimen of pronounced fluorescence is used. It is also possible to use demonstration specimens first.
☞
If compensator
λ
has not been removed from the compartment above the nosepiece after
transmitted-light polarization microscopy, it must be taken out before setting epi-fluorescence.
•
Insert the fluorescence protection shield into the compensator compartment above the nosepiece.
•
Swivel in Plan-Neofluar 20x/0.50 objective.
•
Move the condenser turret to position H, transmitted-light bright field (or phase contrast also), and
then move to the specimen area to be examined.
•
Keep the light path in the incident-light illuminator blocked at first using the shutter plate on reflector
turret (3-28/
1
) or the barrier position of the incident-light filter slider (3-28/
4
).
Summary of Contents for Axioskop 40
Page 1: ...Operating Manual Axioskop 40 Axioskop 40 FL Routine microscope...
Page 14: ...INTRODUCTION Axioskop 40 Carl Zeiss Axioskop 40 FL 0 14 B 40 810 e 12 01...
Page 30: ...INSTRUMENT DESCRIPTION Axioskop 40 Carl Zeiss Axioskop 40 FL 1 16 B 40 810 e 12 01...
Page 77: ...Axioskop 40 OPERATION Axioskop 40 FL Carl Zeiss B 40 810 e 12 01 3 3...
Page 128: ...OPERATION Axioskop 40 Carl Zeiss Axioskop 40 FL 3 54 B 40 810 e 12 01...
Page 146: ...APPENDIX Axioskop 40 Carl Zeiss Axioskop 40 FL A 8 B 40 810 e 12 01...
