This section provides overview, applications, and principles of fluorescence microscopes. Also, please take a look at the list of 13 fluorescence microscope manufacturers and their company rankings.
A fluorescence microscope is a device that uses laser light, super high-pressure mercury vapor lamps, or xenon lamps as a light source to observe the fluorescence of fluorescent substances in an object. In ordinary optical microscopes, visible light such as halogen lamps are used as a light source to irradiate an object and observe reflected light and transmitted light.
Fluorescence microscopy is a type of microscopy that mainly targets biological tissues and cells labeled with fluorescent substances. Since the resolution of a microscope depends on the wavelength of light used, fluorescence microscope that use light with short wavelengths have superior spatial and temporal resolution.
Therefore, highly quantitative information can be obtained. Fluorescence microscope is becoming more and more important as they are used in confocal laser microscopes and multiphoton microscopes, which are becoming more sophisticated.
Fluorescence microscopes are mainly used for bioimaging. The specific targets are cells and tissues, which can be observed while alive. To label an object with fluorescence, a combination of the following techniques are used:
These technologies enable us to observe the localization of target proteins and expressed genes. In addition, drugs and proteins that emit fluorescence in response to specific substances have been developed, making it possible to visualize neural activity and intracellular dynamics of substances.
In recent years, the advent of CRISPR technology has made the creation of genetically modified organisms much easier, and the scope of its application is rapidly expanding.
A fluorescence microscopes are a device that observes fluorescence. Fluorescence is emitted when a fluorescent substance absorbs specific light as energy (excitation light) and releases the energy again.
Exposure to excitation light causes rapid emission of light. The wavelength of fluorescence is longer than the wavelength of the excitation light, and these wavelengths vary with the fluorescent material. In order to observe specific fluorescence, a fluorescence microscopes has a filter unit consisting of the following
By changing or combining filter units, various fluorescent materials can be observed from the same specimen.
The "resolution" of a microscope means the minimum distance at which it is possible to distinguish two close points from different positions. Microscopes use lenses to magnify and observe objects, and in principle, it is possible to increase the magnification infinitely by combining lenses.
However, in the case of optical microscopes, which use light to observe samples, the limit of resolution is approximately half the wavelength of light due to diffraction, which is a characteristic of light. This was considered the theoretical limit of microscope resolution, but a technology was developed that broke through this limit, and the developer was awarded the Nobel Prize in Chemistry in 2014.
The technique is called "super-resolution microscopy". Until the development of super-resolution microscopy, the resolution limit of fluorescence microscopy was approximately 250 nm, but with super-resolution microscopy, the resolution can be as high as 15 to 100 nm, which is close to that of electron microscopy. Super-resolution microscopy achieves high resolution by using various techniques to avoid the limiting factors of resolution.
Super-resolution microscopy methods that have dramatically improved resolution and won the Nobel Prize in Chemistry include "PALM", "STED. PALM" and "STED". They have achieved breakthroughs in fluorescence microscopy resolution by utilizing special optics and special dyes. Super-resolution microscopes using various other technologies have been produced and are being commercialized by various companies.
The advantage of fluorescence microscopy is that it enables detailed observation of molecular behavior and cell structure as visual information. By using the appropriate fluorescence microscopes for a given purpose, an object can be observed with high temporal and spatial resolution.
It is also possible to observe an object using multiple dyes. For example, when two different proteins are labeled with red and green fluorescent substances and observed, a yellow area indicates that these two proteins may exist in the same location in the cell.
Various fluorescent materials and fluorescence microscopes has been developed for different purposes and applications, and are becoming increasingly important in life science and clinical research.
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