OPERATION
Axioskop 2 / Axioskop 2
MOT
Illumination and contrasting techniques
B 40-075 e 06/98
3-39
3.3.6
Setting of epi-fluorescence
(1)
General principle
The epi-fluorescence technique permits high contrast images of fluorescent substances 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), which only allows the long-wave emission from the specimen to be
transmitted.
Excitation and barrier filters, which are both positioned in the FL reflector module together with the
relevant dichroic beam splitter, must be perfectly matched.
(2)
Axioskop 2 (manual) and Axioskop 2
MOT
configuration
−
Recommended objectives: Plan-Neofluar or Fluar (UV-excitation)
−
FL reflector module and shutter module A in reflector turret
−
HBO 103 or HBO 50 mercury vapor short-arc lamp for incident-light illumination
−
HAL 100 halogen illuminator for transmitted-light illumination
☞
Before the epi-fluorescence technique is applied, it is absolutely necessary to adjust the
mercury vapor short-arc lamp in accordance with section 2.1.11 by using the adjusting aid. If
required, re-adjustment must be performed depending on the operation time.
(3)
Setting of epi-fluorescence on the Axioskop 2 (manual) and Axioskop 2
MOT
The first epi-fluorescence setting is considerably facilitated if the Plan-Neofluar objective 20x/0.50 and a
specimen featuring pronounced fluorescence is used. It is also possible to use demonstration specimens
first.
☞
If the
λ
-compensator (3-25/
5
) has not been removed from the compartment above the
nosepiece after transmitted-light DIC microscopy, it must be taken out before setting epi-
fluorescence.
•
Switch on the HAL100 halogen illuminator.
•
Swing in Plan-Neofluar 20x/0.50 objective.
•
Move condenser turret to position H, transmitted-light brightfield (or also phase contrast), and then
move to the specimen area to be examined.
•
Keep light path in the incident-light illuminator blocked at first using shutter module A on the
reflector turret (3-26/
1
) or the barrier position of the incident-light filter slider (3-26/
5
).
Summary of Contents for Axioskop 2
Page 1: ...Axioskop 2 Routine Microscope Operating Manual...
Page 14: ...INTRODUCTION Axioskop 2 Axioskop 2 MOT 0 14 B 40 075 e 06 98...
Page 120: ...OPERATION Axioskop 2 Axioskop 2 MOT 3 54 B 40 075 e 06 98...
Page 122: ...CARE MAINTENANCE TROUBLESHOOTING AND SERVICE Axioskop 2 Axioskop 2 MOT 4 2 B 40 075 e 06 98...
Page 132: ...CARE MAINTENANCE TROUBLESHOOTING AND SERVICE Axioskop 2 Axioskop 2 MOT 4 12 B 40 075 e 06 98...
Page 134: ...ANNEX Axioskop 2 Axioskop 2 MOT A 2 B 40 075 e 06 98...
