The differences between fluorescence microscope and optical microscope

The differences between fluorescence microscope and optical microscope 



Fluorescence microscope and optical microscope is different, it is not the ordinary lighting on the samples, but the use of certain wavelengths of light (typically ultraviolet light, blue light excitation fluorescence microscope) within the specimen, the emission fluorescence, so light, the fluorescence microscope is not the role of the direct lighting, but as a fluorescence material excited specimens of energy. We can observe the specimen, not due to lighting, fluorescent light fluorescent material excited phenomenon showing absorption but within the specimen after. Therefore, the characteristics of fluorescence microscopy, especially its light source can supply a large number of specific wavelength range of the excitation light, so that by the fluorescent material inspection within the specimen to obtain the necessary intensity of luminescence. At the same time, fluorescence microscopy must have the corresponding filter system. Fluorescence microscopy is a basic tool for immunofluorescence histochemistry. It is composed of ultra high pressure light source, filter system (including excitation and suppression of the filter plate), the main components of optical system and camera system, is to use a certain wavelength to excite the samples emit fluorescence.

1 Fluorescence excitation way: according to the wavelength of the light is divided into UV excitation method (using ultraviolet illumination method) and BV excitation method (using the blue violet) two. UV excitation method is near ultraviolet light with shorter than 400nm excitation. The method does not exist the visible luminescence, fluorescence and fluorescence was observed to render the dye inherent, easy to discriminate specimens on the specific fluorescence and background tissue autofluorescence.

 2.The BV excitation method: 404nm, 434nm as the center from the UV to blue light excitation. The method uses blue light irradiation specimen, so the fluorescence observation system cut-off filter must be used to completely block the blue and green, Huang Yingguang fully through the required filter. For fluorescent pigment fluorescent antibody method. The maximum emission wavelength is close to the maximum absorption wavelength of the excitation light and fluorescence, so BV excitation filter method used must use a sharp cut-off filter. The method can be used as excitation light blue light, so the fluorescent pigment absorption efficiency is high, can get the image brighter. The disadvantage is the fluorescence below 500nm and above 500nm couldn't see, so that the image color yellow. The fluorescent antibody method, mostly to the fluorescent pigment specific color to determine its specificity, so the discussion of specific subtle, the BV excitation method often has great influence.

Post time: 2014-07-03

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