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(1) Fluorescence microscopes are generally divided into two types, transmission and epi-illumination.
a. Transmissive: the excitation light comes from below the object to be inspected. The condenser is a dark field condenser, so that the excitation light does not enter the objective lens, but the fluorescence enters the objective lens. It is bright at low magnification and dark at high magnification. It is more difficult to operate during oil immersion and mid-adjustment. Especially the low magnification illumination range is difficult to determine, but it can get a very dark field of view background. The transmission type is not used for non-transparent objects.
b. Epi-illumination: Transmittance is almost eliminated nowadays. Most new fluorescence microscopes are epi-illumination. The light source comes from above the object to be inspected and has a beam splitter in the optical path, so it is suitable for both transparent and opaque objects. Because the objective lens acts as a condenser lens, it is not only easy to operate, but also can achieve uniform illumination of the entire field of view from low to high power.
(2) Precautions for fluoroscopy
a. If the excitation light is irradiated for a long time, the phenomenon of fluorescence attenuation and quenching will occur, so the observation time should be shortened as much as possible. When not observing temporarily, the excitation light should be covered with a baffle.
b. Use "no fluorescent oil" for oil lens observation.
c. Fluorescence is almost always weak and should be performed in a dark room.
d. The power supply is best equipped with a voltage stabilizer, otherwise the unstable voltage will not only reduce the life of the mercury lamp, but also affect the effect of microscopic examination.
At present, many emerging biological research fields are applied to fluorescence microscopy, such as gene in situ hybridization (FISH) and so on.
Fluorescence microscope is to irradiate the test object stained with fluorescein with short-wavelength light, make it excited and produce long-wavelength fluorescence, then observe. Fluorescence microscopes are widely used in biology, medicine and other fields.
Fluorescence CCDs usually use cooled CCDs, because cooled CCDs can reduce background noise and are good for low fluorescence signal acquisition, especially like single molecule imaging. It is not necessary to cool the CCD to collect strong fluorescent signals. Black and white cameras can be used. Color cameras are not suitable for fluorescent collection because of their low sensitivity and low resolution. Unless the fluorescent signal is very high and the resolution requirements are not high, it is suitable for color. camera.