What are the differences between fluorescence microscopy and inverted microscopy
Microscope is an important instrument in cell culture and related derivative experiments. At present, there are various types of microscopes on the market, and choosing a microscope that meets the needs and is suitable is a challenge. Below, we will introduce the principles of inverted microscopes and fluorescence microscopes for everyone to choose from.
The inverted microscope, like a regular microscope, mainly consists of three parts: mechanical part, lighting part, and optical part.
The composition of an inverted microscope is the same as that of a regular upright microscope, except that the objective lens and lighting system are reversed, with the former under the stage and the latter above the stage.
This structure significantly expands the effective distance between the lighting spotlight system and the stage, making it easier to place thicker observation tools such as culture dishes and cell culture bottles (of course, glass slides can also be used), while the working distance between the objective lens and the material does not need to be very large.
Inverted microscope is used by medical and health institutions, universities, and research institutes for observing microorganisms, cells, bacteria, tissue cultures, suspensions, sediments, etc. It can continuously observe the process of cell and bacterial proliferation and division in culture medium, and can capture any form of this process.
Widely used in fields such as cytology, parasitology, oncology, immunology, genetic engineering, industrial microbiology, and botany.
Fluorescence microscopy is used to study the absorption, transportation, distribution, and localization of substances within cells.
For the tested object, there are two ways to generate fluorescence: spontaneous fluorescence, which is directly emitted by ultraviolet irradiation; Secondary fluorescence occurs when the observed object is treated with fluorescent dyes and exposed to ultraviolet light before emitting fluorescence.
Some substances in cells, such as chlorophyll, produce spontaneous fluorescence after being exposed to ultraviolet radiation; Some substances themselves may not emit fluorescence, but if stained with fluorescent dyes or fluorescent antibodies, they can also emit secondary fluorescence under ultraviolet radiation.
A fluorescence microscope uses a high luminous efficiency point light source to emit a certain wavelength of light (UV 365nm or UV blue 420nm) through a color filtering system as excitation light, which excites the fluorescent substances in the sample to emit various colors of fluorescence. After that, it is observed through magnification of the objective lens and eyepiece.
In this way, even with weak fluorescence, it is easily recognizable and highly sensitive under strong contrasting backgrounds. It is mainly used for studying cell structure, function, and chemical composition.
