Difference between ordinary optical microscope and fluorescence-based microscope
Optical microscope: It is an optical instrument that uses optical principles to magnify and image tiny objects that cannot be distinguished by the human eye, so that people can extract detailed structural information.
Fluorescence microscope: A fluorescence microscope uses ultraviolet light as a light source to illuminate the object being inspected to make it emit fluorescence, and then observe the shape and location of the object under the microscope. Fluorescence microscopy is used to study the absorption and transport of substances within cells, as well as the distribution and positioning of chemical substances. Some substances in cells, such as chlorophyll, can fluoresce after being irradiated by ultraviolet rays; some substances themselves cannot fluoresce, but if they are stained with fluorescent dyes or fluorescent antibodies, they can fluoresce after irradiation with ultraviolet rays. Fluorescence microscopy is the One of the tools for qualitative and quantitative research on such substances.
Optical microscopes generally consist of a stage, a spotlight illumination system, an objective lens, an eyepiece and a focusing mechanism. By changing the lighting method, the spotlight lighting system can obtain different observation methods such as dark object points on a bright background (called bright field lighting) or bright object points on a dark background (called dark field lighting), so that it can be used in different situations. to better discover and observe fine structures.
The differences between fluorescence microscope and ordinary microscope are as follows:
1. The lighting method is usually epi-illumination, that is, the light source is projected on the sample through the objective lens;
2. The light source is ultraviolet light, with shorter wavelength and higher resolution than ordinary microscopes;
3. There are two special filters, the one in front of the light source is used to filter visible light, and the one between the eyepiece and objective lens is used to filter ultraviolet rays to protect human eyes.
Fluorescence microscopy is also a type of optical microscope. The main difference is the different excitation wavelengths. This determines the differences in structure and usage between fluorescence microscopes and ordinary optical microscopes.
Fluorescence microscopy is an essential tool in immunofluorescence cytochemistry. It is composed of main components such as light source, filter plate system and optical system. It uses light of a certain wavelength to excite the specimen to emit fluorescence, and then magnifies the fluorescence image of the specimen through the objective lens and eyepiece system.
