Zeiss Axiovert 200 Operating Manual

Also see for Axiovert 135M: Operating manual Instructions for use

Contents

OPERATIONCarl Zeiss Illumination and contrasting techniques Axiovert 2003-40 B 40-080 e 03/013.3.5 Setting of fluorescence contrast in reflected light3.3.5.1 General principleThe epi-fluorescence technique permits high-contrast images of fluorescent substances in typicalfluorescence colors. In the epi-fluorescence microscope, the light generated by a high-performanceilluminator reaches the excitation filter via a heat-reflecting filter. The filtered, short-wave excitationemission is reflected from a dichroic beam splitter and is focused on the specimen via the objective. Thespecimen absorbs the short-wave emission and then emits long wave fluorescence (STOKE's law) whichis now gathered by the objective and transmitted by the dichroic beam splitter. Finally, the beams pass abarrier filter which only allows the long-wave emission from the specimen to be transmitted.Excitation and barrier filters, which are positioned in the FL reflector module together with theappropriate dichroic beam splitter, must be perfectly matched.3.3.5.2 Configuration of the Axiovert 200 (manual) and Axiovert 200 M− Recommended objectives: brightfield objectives− FL reflector module in the reflector turret− N HBO 103 fluorescence illuminator or HBO 50 for reflected-light illumination− HAL 100 halogen illuminator for transmitted-light illumination Before the epi-fluorescence technique is applied, it is absolutely necessary to adjust themercury vapor short-arc lamp in accordance with sections 2.12.2 through 2.12.4 by using theadjusting aid. If required, re-adjustment must be performed depending on the operation time.3.3.5.3 Setting of epi-fluorescence on the Axiovert 200 (manual) and Axiovert 200 MThe first epi-fluorescence setting is considerably facilitated if the Plan-Neofluar objective 20x/0.50 and aspecimen featuring pronounced fluorescence are used. It is also possible to use demonstration specimensfirst.• Switch on the HAL 100.• Swing in suitable objective, e.g. Plan-Neofluar 20x/0.50, via the nosepiece (3-24/4).• Move condenser turret to position H, transmitted-light brightfield (or also phase contrast), and thenmove to the specimen area to be examined.• Use focusing drive for focusing.• Keep light path in the reflected-light part blocked at first using the fluorescence shutter by pressingthe FL on / off key (3-24/6).