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Mysterious fluorescent colors: Do you know how to detect biomolecules using different wavelengths of light?
In today's biomedical research, fluorescence imaging technology is like a key that opens the door to tightly closed biological processes. This non-invasive technology allows us to observe biological processes in living organisms and thus understand the mysteries of life. Using a variety of methods including microscopy, imaging probes and spectroscopy, scientists are able to capture dynamic changes within cells, such as gene expression and protein interactions.
Fluorescence is a form of luminescence in which a substance emits light of a specific wavelength after absorbing electromagnetic radiation. Molecules that are able to re-emit light after absorbing it are called fluorescent molecules.
Fluorescence Mechanism
When a molecule absorbs light, its energy is briefly boosted to a higher excited state. When it returns to its ground state, it emits fluorescent light that can be detected. This emitted light has a certain wavelength, and this wavelength is what we need to know before the experiment to ensure that the measurement device can correctly detect the generation of light.
Fluorescent dyes and proteins
Fluorescent dyes and fluorescent proteins each have their own advantages and disadvantages. Fluorescent dyes do not require maturation time and generally have higher photostability and brightness than fluorescent proteins. For example, green fluorescent protein (GFP) fluoresces green when illuminated by light in the ultraviolet range and is an excellent reporter molecule for observing protein binding and gene expression.
Imaging range and system
Fluorescence imaging is typically performed using a charge-coupled device (CCD), which can accurately detect and image light in the 300-800 nanometer range. The widespread use of this technology allows us to capture biological processes that cannot be seen by the naked eye in experiments.
The intensity of the fluorescence signal exhibits relatively linear behavior with the number of fluorescent molecules, which is a major advantage of fluorescence imaging.
Applications
Fluorescence imaging plays an important role in many biological applications. For example, in PCR technology, SYBR green dye is widely used to visualize DNA. In cancer surgery, fluorescence imaging can assist surgeons in precisely locating cancerous tissue during tumor removal.
Types of Microscopes
Different microscopy techniques can alter the visualization and contrast of the image. Total internal reflection fluorescence microscopy is a technique that uses Leydig waves to selectively observe the fluorescence of single molecules, while beam fluorescence microscopy illuminates the sample at a perpendicular angle to emphasize specific layers.
Advantages and Disadvantages The advantage of fluorescence imaging technology is that it is non-invasive, so it can be performed in living bodies without damaging the skin. However, its limitations cannot be ignored, such as fluorescence quenching and the influence of environmental factors on fluorescence efficiency, which may interfere with the imaging results.Future Directions
Scientists continue to work on developing more effective fluorescent proteins by genetically engineering them to alter their fluorescent properties in order to improve the capabilities of imaging probes. In addition, fluorescence resonance energy transfer (FRET) and fluorescence correlation spectroscopy (FCS) techniques have the potential to further enhance the sensitivity and range of fluorescence imaging, bringing more possibilities to biomedical research.
On the whole, fluorescence imaging technology is not only an important tool for exploring the mysteries of life, but also opens up new directions for future biomedical research. In the future, will we be able to see more precise biological processes and deeper life phenomena?
