Zeiss Axiolab A Especificaciones descargar pdf (Pagina 105)

OPERATION

Axio Lab.A1 Lighting and contrasting method in reflected light Carl Zeiss

04/2013 430037-7144-001 105

4.2.4 Adjusting reflected light fluorescence

(1) General principle

The reflected light fluorescence method is used to show fluorescent substances in typical fluorescence

colors in high contrast. The light originating from a high-performance illuminator in a reflected light

fluorescence microscope passes through a heat protection filter to an excitation filter (bandpass). The

filtered, short-wavelength excitation beam is reflected by a dichroic beam splitter and is focused on the

specimen above the objective. The specimen absorbs the short-wave radiation before emitting longer-

wave fluorescence radiation (Stokes’ Law). This radiation is then captured from the image side by the

objective and passes through the dichroic beam separator. Finally, the beams pass through a band

elimination filter (longpass/bandpass) which only permits the long-wave radiation emitted by the

specimen to pass through.

The spectra of the excitation and the band-elimination filters must match very closely. They must be

inserted in a reflector module FL P&C together with the respective dichroic beam splitter.

Only powerful LED are supplied as FL excitation light sources in the Axio Lab.A1 program.

(2) Instrumentation

Observations in reflected light fluorescence can only be made on Axio Lab.A1 microscopes for reflected

light and reflected light fluorescence.

 Recommended objectives: EC Plan-Neofluar or Fluar (UV excitation)

 LED modules for FL excitation (maximum two configurable)

 Reflector modules FL P&C equipped with respective filter sets

 Fluorescence protection shield

(3) Adjusting reflected light fluorescence

The adjustment of reflected light fluorescence is facilitated by starting with an objective of average

magnification, e.g. EC Plan-Neofluar 20x/0.50, and a specimen of high fluorescence. Demonstration

samples can also be used for the start-up.

If the compensator O used for the transmitte

d light polarization method is still in its

compartment above the nosepiece, it must be removed before adjusting reflected light

fluorescence.

x Slide the fluorescence protection shield (Fig. 4-20/8) into the compensator compartment above the

nosepiece.

x Swivel in the 20x/0.50 EC Plan-Neofluar objective on the nosepiece (Fig. 4-20/4).

x Initially set the FL/TL toggle switch (Fig. 4-20/2) to position TL (transmitted light).

x If necessary, turn the condenser turret (Fig. 4-20/7) to position H transmitted light-brightfield (or phase

contrast if using a Ph objective) and seek the specimen detail to be examined.

x Adjust the light intensity by turning the regulator (Fig. 4-20/5) and focus (Fig. 4-20/6).

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Zeiss Axiolab A Especificaciones Pagina 105