OPERATION
Carl Zeiss
Illumination and Contrast Techniques in Reflected Light
Axio Vert.A1
94
431030-7044-001
05/2012
4.12.5
Reflected Light Fluorescence
The reflected light fluorescence method allows fluorescent substances to be presented in a high-contrast
manner and in typical fluorescence colors. In a reflected light fluorescence microscope, the light produced
by a high-performance illuminator reaches the excitation filter through a heat protection filter. The
filtered short-wavelength excitation radiation is reflected by a dichroic beam splitter and then focused on
the specimen via the objective. The specimen absorbs the short-wavelength radiation and subsequently
emits fluorescence radiation of a longer wavelength (Stoke's Law), which is picked up as an image by the
objective and transmitted by the dichroic beam splitter. The rays eventually pass through a barrier filter,
which allows only the long-wavelength radiation emitted by the specimen to go through.
The excitation and barrier filters need to be very accurately matched spectrally and are located in an FL
P&C reflector module, together with the corresponding dichroic beam splitter.
Requirements
−
Stand Axio Vert.A1 FL or Axio Vert.A1 with HBO / HXP illuminator
or
−
Axio Vert.A1 FL-LED stand with LED modules mounted
−
The microscope must have been properly put into operation and switched on, as described in
Section 3.
−
Bright field objectives
−
FL P&C reflector module in the reflector turret
−
Illuminators for fluorescence (HBO 50, HBO 100, Colibri 2, HXP 120 C)
−
Prior to their use, the HBO 50 or HBO 100 illuminators need to be adjusted and warmed up for about
15 minutes, as described in Section 3.1.11.2 and 3.1.11.3, respectively.
−
HAL illuminator or LED illuminator for transmitted light illumination
Setting Fluorescence
•
Select the area of the specimen in transmitted light bright field or phase contrast. To this end, switch
the reflector turret (Fig. 4-17/
4
) to free passage and set the position for bright field or phase contrast
on the condenser (Fig. 4-17/
2
).
•
Use the power supply unit (Fig. 4-17/
7
) to turn on the fluorescence illumination, e.g., the HBO 50
illuminator (Fig. 4-17/
3
).
•
Use the reflected light shutter (Fig. 2-7/
25
) to keep the light path temporarily blocked.
•
Once you have selected the area of the specimen, use the
on/off
function (Fig. 4-17/
5
and
6
) to
switch off the transmitted light illumination (Fig. 4-17/
1
).
When you employ the Axio Vert.A1 FL-LED stand, the
TL
/
RL
switch (Fig. 2-8/
20
) lets you shift
directly between transmitted light and reflected light.
•
Using the reflector turret (Fig. 4-17/
4
), rotate in the desired filter set and unblock the light path with
the reflected light shutter (Fig. 2-7/
25
).
•
Close the luminous field diaphragm (Fig. 4-17/
10
) until it is visible in the image. Using the centering
screws, move the diaphragm into the center position and open it all the way to the edge of the field
of view.
Содержание Axio Vert.A1
Страница 1: ...Operating Manual Axio Vert A1 Inverted Microscope ...
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