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From light source to imaging: What is the technological miracle behind fluorescence microscopy?
Fluorescence microscopy is a type of optical microscope that uses fluorescence to study the properties of organic or inorganic substances. The microscope design can be simple, such as an epifluorescence microscope, or more complex, such as a confocal microscope, which uses optical sectioning techniques to obtain better resolution of fluorescence images. This technology is based on shining light of a specific wavelength onto a sample, with fluorescent substances in the sample absorbing this light and re-emitting light at longer wavelengths, creating the fluorescent image we see.
The main components of a fluorescence microscope include a light source, excitation filter, grating mirror and emission filter. These components are carefully selected to match the characteristics of the fluorescent markers used.
The importance of light source
Fluorescence microscopes have extremely high requirements for lighting. Common light sources include xenon arc lamps and mercury lamps. However, with the advancement of technology, lasers, high-power LEDs and supercontinuum light sources have also begun to be used. Lasers are commonly used in more sophisticated microscopy techniques, while xenon and mercury lamps are commonly used for excitation filtering in widefield fluorescence microscopy.
Sample preparation and labeling technology
Samples must be appropriately fluorescently labeled before they can be observed with a fluorescence microscope. These methods include labeling with fluorescent dyes or expressing fluorescent proteins in biological samples. In this way, scientists can precisely detect the distribution of proteins or other molecules in cells.
Biofluorescent dyes are designed to cover a variety of biomolecules, and these dyes can specifically label specific structures inside cells.
Immunofluorescence technology
Immunofluorescence technology uses the highly specific binding between antibodies and antigens to label specific proteins or other molecules in cells. This technology allows researchers to clearly observe microtubules or other structures in cells. This highly precise labeling method makes fluorescence microscopy play an important role in biomedical research.
Limitations and Challenges
Although fluorescence microscopy has demonstrated great capabilities in the field of biology, it also faces some challenges. For example, fluorescent substances will photobleach under long-term exposure, which will affect the effectiveness of observation. In addition, fluorescent molecules tend to produce reactive chemicals when exposed to light, further exacerbating the potential damage to the specimen being observed.
To overcome these limitations, researchers are working to develop more stable fluorescent reagents and sampling techniques.
The rise of super-analytic technology
The wave nature of light limits the resolution of microscopes. However, with the advancement of science and technology, many new technologies have emerged, such as STED microscopes and multi-photon stimulation technology, which allow scientists to surpass the traditional diffraction limit and obtain Higher details and resolution.
With the development of fluorescence microscopy technology, we have not only seen the huge role this technology plays in the field of life sciences, but also witnessed its potential in other scientific fields. In the process of exploring the microscopic world, how will these technological miracles affect future scientific exploration?
