Principle and application of inverted fluorescence microscope

2022-12-25 12:05:30
Inverted fluorescence microscope introduction:
The inverted fluorescence microscope is composed of a fluorescent accessory and an inverted microscope. It is mainly used for fluorescence and phase contrast observation of living tissues such as cells. Inverted microscope is used for microscopic observation of tissue culture, cell culture, plankton, environmental protection, food inspection, etc. in the fields of biology and medicine. Since these living objects are placed in a culture dish (or a culture flask), the objective lens and the condensing lens of the inverted microscope are required to have a long working distance, and the object to be inspected in the culture dish can be directly observed and studied. Therefore, the positions of the objective lens, the condensing mirror, and the light source are reversed, so it is called an "inverted microscope." Inverted microscopes are mostly used for colorless and transparent living observation. A set of fluorescent accessories is added on the basis of the inverted microscope: laser excitation block, fluorescent light source, fluorescent illuminator, and excitation block switching device, so that inverted fluorescence observation can be performed.

Inverted fluorescence microscopy is a new type of fluorescence microscope developed in modern times. It is characterized by excitation light falling from the objective lens to the surface of the specimen, that is, using the same objective lens as an illumination concentrator and a fluorescent collection objective lens. A two-color beam splitter is added to the optical path, which is at an angle of 45° to the uranium. The excitation light is reflected into the objective lens and collected on the sample. The fluorescence generated by the sample is reflected by the surface of the objective lens and the surface of the cover glass. The excitation light enters the objective lens at the same time, and returns to the two-color beam splitter to separate the excitation light from the fluorescence, and the residual excitation light is absorbed by the blocking filter. For example, a combination of different excitation filter/two-color beam splitter/blocking filter inserts can meet the needs of different fluorescent reaction products. The advantage of such a fluorescence microscope is that the field of view illumination is uniform, the imaging is clear, and the larger the magnification, the stronger the fluorescence.

Inverted microscope application:
Inverted microscopes have the characteristics of microscopic observation in culture flasks or petri dishes; transparent living bodies without staining can be observed; epifluorescence microscopy is suitable for fluorescence microscopy. The instrument is especially suitable for microscopic research on living cells and tissues, fluids, sediments, etc. It is an ideal instrument for research work in biology, cytology, oncology, genetics, immunology and so on. It can be used by scientific research, universities, medical care, epidemic prevention, agriculture and animal husbandry. The inverted fluorescence microscope consists of an inverted microscope and an epifluorescence microscope. The instrument is equipped with a long working distance flat field achromatic objective lens, a large field of view eyepiece, and binocular observation. The inverted microscope is also equipped with a special or long working distance concentrating mirror, and is equipped with a phase contrast device and a long working distance flat field phase contrast objective lens.

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