Fluorescent microscopy
| Fluorescent microscopy | |
|---|---|
 | |
| Technology Details | |
| Other Names | Epi-fluorescense |
| Technology | Metrology |
| Equipment | List of fluorescent microscopy equipment |
| Materials | Biological samples that have been fluorescent labeled |
Fluorescent microscopy is an optical microscope technique widely used in Biomedical Sciences. luminescence family of processes in which susceptible molecules emit light from electronically excited states created by either a physical, mechanical, or chemical mechanism when illuminated with a powerful light source such as mercury or xenon arc (burner) lamps. Fluorescence microscopy is capable of imaging the distribution of a single molecular species based solely on the properties of fluorescence emission.
Contents
Equipment
Fluorescent microscopes are inverted microscopes, which allows a close approach to the specimen usually placed on a glass slide. The microscope must have a light source powerful enough to excite the fluorescent process in the tagged cells. In addition it is equipped with optical filters that allow only a narrow range of wavelength to selectively excite different photo-luminescent molecules,
Olympus BX 51
Method of operation
Fluorescense:
A Chromophore is a naturally occurring or synthetic pigment with characteristic optical absorption, usually containing a combination of alternating single and double bonds or a high degree of cyclic aromatic or heterocyclic conjugation. In optical microscopy, chromophores often refer to classical dyes, such as Eosin, Safranin, or Hematoxylin, utilized to stain tissues for brightfield observation. Fluorescent probes are also classified as chromophores because they exhibit strong absorption spectra in the ultraviolet, visible, and/or near infrared regions.
| Wavelength Range (nm) | Color |
|---|---|
| 340-400 | Ultraviolet |
| 400-430 | Violet |
| 430-465 | Blue |
| 465-500 | Cyan |
| 500-550 | Green |
| 550-580 | Yellow |
| 580-620 | Orange |
| 620-700 | Red |
| Over 700 | Near Infrared |
Parameters
Here we describe what parameters are of importance in this technology (e.g. power, temperature...). May not be relevant to some technologies.
Applications
Fluorescent Microscopy is the most common technique used in biological sciences to study live cells and cellular processes while recording image data. Basically, cells are tagged with a fluorophore, when this molecule is excited it briefly releases of a particular energy which allows their detection and identification. A fluorescent tag is a molecule that is attached chemically to a a biomolecule such as a protein, antibody, or amino acid to aid in the labeling and detection. The fluorophore selectively binds, chemically or biologically, to a specific region or functional group on the target molecule. Various labeling techniques such as enzymatic labeling, protein labeling, and genetic labeling are widely utilized. The most commonly labelled molecules are antibodies, proteins, amino acids and peptides which are then used as specific probes for detection of a particular target in the cell. The introduction of genetically-encoded fluorescent protein fusions as a localization marker in living cells has revolutionized the field of cell biology lately.
Materials
Optional description of materials that can be processed by technology. I think the best example of where this comes in handy would be with LPCVD describing the difference between HTO and LTO.
See also
Other related wiki pages
References
Further reading
- video [1]
