Proper Use of an Inverted Fluorescence Microscope
Inverted fluorescence microscope is a multifunctional microscope used in the high-end field of biological science research. Its structure is complex and expensive. Therefore, in order for medical graduate students to correctly master the operation and use specifications of inverted fluorescence microscope, teachers must first provide a course on the basic knowledge of inverted fluorescence microscope before demonstrating the actual operation of inverted fluorescence microscope, introducing the structure, principle, function and basic usage methods of inverted fluorescence microscope. Then, in the specific operation, different functions are demonstrated for different specimens, gradually standardizing the operation and use of inverted fluorescence microscope by medical graduate students.
1. Correct use of clear vision observation. One of the functions of inverted fluorescence microscope is bright field observation. Its basic principle is that after the specimen is irradiated with light, different parts absorb light differently to achieve imaging in a bright background. It is mainly suitable for observing specimens such as stained cells and tissue sections that have their own color.
2. Correct use of differential observation. The second function of inverted fluorescence microscope is phase difference observation. Its basic principle is to convert the optical path difference of different parts of the sample into amplitude difference, which is observed through a phase difference objective lens with a phase plate and a condenser lens with a circular aperture. It is mainly suitable for observing live cells or unstained tissue sections. In the process of experimental teaching, in addition to guiding medical graduate students to follow the basic operation methods of phase difference observation correctly, it is more important to guide them to understand and master the specimens using phase difference observation. For them, the most commonly used method in future scientific research is microscopic observation of cultured cells. Firstly, for the selection of cell equipment, it is generally recommended to use glass and plastic culture dishes, plates, and bottles. They are mostly colorless and transparent, and can be directly observed under an inverted fluorescence microscope without the need to discard cell fluid or undergo other treatments. Secondly, during microscopic observation, do not open the lid of the equipment casually to avoid contaminating the cells. Finally, for operators, it is necessary to wear gloves, masks, and white coats, which not only protect their own health but also prevent cell contamination.
