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Ultraviolet (UV) light is non-ionizing radiation, emitted by the sun and some artificial sources. Some lasers, incandescent, halogen, fluorescent, and mercury vapor lamps serve as artificial sources of UV radiation. The light is classified into UVA, UVB, and UVC. The wavelength of UVA is 315-399 nm. The wavelength of UVB is 280-314 nm. Then, the wavelength of UVC is 100-279 nm. Around all of the UV radiation falling on the earth is UVA since UVB and UVC are mostly absorbed by ozone. Continuous exposure to UV light can cause premature aging, sunburn, skin cancer, eye diseases, melanoma, squamous cell cancer, and basal cell cancer.
A UV transilluminator is a typical equipment present in biosciences laboratories for the visualization of target proteins and DNA. The transilluminator utilizes intense UV irradiation to make visible the fluorescent tags used in gels. Ethidium bromide is a commonly used fluorescent marker in the laboratory for visualization under UV transilluminator.
The main usage of a UV transilluminator is in the visualization of protein and DNA polyacrylamide and agarose gels after the process of electrophoresis. It is used for gel electrophoresis, protein fluorescence, and gel fluorescence. It is predominantly used when ethidium bromide is used as a fluorescent tag. It is used to find out the tumor and facilitate medical inspection. It provides researchers with the compact footprint of the gel. The gels are placed directly on the surface of the UV transilluminator. But the wavelength will change depending on the specific application.
When a transilluminator is illuminated at the wavelength of 365 nm, there is no occurrence of photonicking, thus facilitating the usage of UV radiation for extended periods. This can be used for routine imaging in a gel documentation system. A wavelength of 302 nm is useful for fast observation of a single band. Illumination at 245 nm can be used for the recording of gels.
Transilluminators are used for the below mentioned purposes:
For different sizes of gels, the viewing surface area differs. Wavelength also differs as the concentration and purpose of gel imaging change. Some transilluminators have a single wavelength, while some have a facility for dual wavelengths. The dimensions of the benchtop unit also vary as some are very compact, while some are elongated. In addition, some models are stationary, whereas some are hand-held/portable. Some models have an option of variable intensity setting, whereas some are without the choice of intensity setting. Some models have options to choose high, medium, and low levels of intensity of irradiation. Some models have only a high level of setting. The switch on the transilluminator is called triple intensity or dual intensity switch or single intensity switch depending on the option of having wavelengths. Smaller size transilluminators can accommodate midi and mini-sized gels. For highly demanding experiments, stationary ones are chosen.
Since intense UV light is used for visualization of fluorescent markers used in agarose gel electrophoresis, the radiation is perilous to both eyes and skin. Hence, a UV blocking cover is provided with each transilluminator. If the instrument is used without putting the cover or if there is no cover, then it is compulsory to protect the exposed eyes and skin of the operator. Users should be made aware not of the glass door in the transilluminator in cutting the gel.
Protection is done by wearing:
From the above paragraphs, we come to know about the specifications and applications of UV transilluminators. This type of illuminator is cheaper compared to LED light transilluminators. But choosing an ethidium bromide fluorescent tag for UV light illumination compromise on disposal of gel and the safety of the operator.