Confocal laser microscopy and fluorescence microscopy: Differences and Similarities
1. Different principles
1. Fluorescence microscope: It uses ultraviolet light as the light source to irradiate the object under inspection to make it emit fluorescence, and then observe the shape and location of the object under the microscope.
2. Laser confocal microscope: a laser scanning device is installed on the basis of fluorescence microscope imaging, and fluorescent probes are excited by ultraviolet light or visible light.
Two, different characteristics
1. Fluorescence microscope: used to study the absorption, transportation, distribution and localization of substances in cells, etc. Some substances in cells, such as chlorophyll, can fluoresce after being irradiated by ultraviolet rays; although some substances themselves cannot fluoresce, they can also fluoresce after being stained with fluorescent dyes or fluorescent antibodies after being irradiated by ultraviolet light.
2. Laser confocal microscopy: use computer for image processing to obtain fluorescent images of the microstructure inside cells or tissues, and observe physiological signals such as Ca2+, pH value, membrane potential and changes in cell morphology at the subcellular level.
Three, different uses
1. Fluorescence microscope: Fluorescence microscope is the basic tool of immunofluorescence cytochemistry. It is composed of main components such as light source, filter plate system and optical system. It is to use a certain wavelength of light to excite the specimen to emit fluorescence, and to observe the fluorescence image of the specimen by amplifying the objective lens and eyepiece system.
2. Laser confocal microscopy: Laser scanning confocal microscopy has been used in the study of cell morphological localization, three-dimensional structure reorganization, and dynamic change processes, and provides practical research methods such as quantitative fluorescence measurement and quantitative image analysis. Combined with other related biotechnologies, it has been widely used in the fields of molecular and cell biology such as morphology, physiology, immunology, and genetics.
