Technical parameters and usage skills of fluorescence microscope
Technical parameters of fluorescence microscope 1. Wide angle eyepiece 2. Achromatic objective lens 3. Four hole objective lens converter 4. Falling fluorescence device Blue (B) Green (G) excitation system 100W mercury lamp 5. Coaxial coarse micro focusing mechanism: focusing range 15mm Micro grid value 0.002mm 6. Double layer mechanical workbench longitudinal movement range: 70mm Lateral movement range: 50mm
Technical parameters of fluorescence microscope
1. Wide angle eyepiece
2. Achromatic objective lens
3. Four-hole objective lens converter
4. Falling Fluorescence Device Blue (B) Green (G) Excitation System 100W Mercury Lamp
Coaxial coarse micro focusing mechanism: focusing range: 15mm, micro grid value: 0.002mm
6. Double layer mechanical workbench
Longitudinal movement range: 70 mm Lateral movement range: 50 mm
Tips and methods for using fluorescence microscopy
（1） Slide
The thickness of the slide should be between 0.8 and 1.2 mm. If the slide is too thick, on the one hand, it absorbs too much light, and on the other hand, it cannot cause excitation light to gather on the specimen. The slide must be smooth and clean, with a uniform thickness, and without significant spontaneous fluorescence. Sometimes quartz glass slides are needed.
（2） Cover glass
The thickness of the cover glass is about 0.17mm, and it is smooth and clean. In order to enhance excitation, interference cover glass can also be used. This is a special cover glass coated with several layers of substances (such as magnesium fluoride) that have different interference effects on different wavelengths of light. It can allow fluorescence to pass smoothly, while reflecting excitation, which can excite the sample.
（3） Specimen
Tissue sections or other specimens should not be too thick. For example, if they are too thick, most of the excitation light is consumed in the lower portion of the specimen, while the upper portion directly observed by the objective lens is not sufficiently excited. In addition, cell overlap or impurity masking can affect judgment.
（4） Mounting agent
Glycerin is commonly used as a mounting agent, and it must be free of spontaneous fluorescence, colorless and transparent. The brightness of the fluorescence is brighter at pH 8.5 to 9.5, and it is not easy to fade quickly. Therefore, an equivalent mixture of glycerol and 0.5 mol/l carbonate buffer solution with a pH of 9.0 to 9.5 is commonly used as a mounting agent.
（5） Mirror oil
Generally, when using a dark field fluorescence microscope and oil microscope to observe specimens, it is necessary to use lens oil. It is best to use a special non fluorescent lens oil, which can also be replaced by the aforementioned glycerol. Liquid paraffin can also be used, but the refractive index is low, which has a slight impact on image quality.
