Kaede | Kaede
Key Words: confocal, Fluorescence, fluorophore, live-cell imaging
Definition:Kaede is a green fluorescing protein that can be irreversibly converted to red on irradiation with UV light
TECHNOLOGY:
Kaede is a protein that was isolated from the stony coral Trachyphyllia geoffroyi. The protein contains a tripeptide, His 62-Tyr 63-Gly 64. This is a green chromophore that is photoconvertible to red following UV irradiation (350-410 nm light). Maximal illumination results in a 2000-fold increase in the ratio of red-to-green fluorescence. Kaede has the benefit that the excitation light used to elicit green fluorescence does not induce photoconversion. Photoconversion can be carried out selectively with appropriate targeting of UV excitation light.
APPLICATIONS:
As with the GFPs, Kaede can be genetically encoded in cells and is easily detectable by standard fluorescence microscopy techniques. Kaede has wide application in molecular and cell biology and provides a simple technique for marking individual cells, organelles and proteins in live cell imaging applications.
MICROSCOPE CONFIGURATION:
Kaede can be visualized on any upright microscope or inverted microscope with epi-fluorescence capability or stereo microscope with stereo fluorescence illuminator (also known as GFP-illuminator). The full potential for imaging live cell dynamics using Kaede is realized in confocal imaging.
RECOMMENDED SYSTEM:
For full flexibility in the imaging of kaede in living cells, the Ti series microscopes configured with wide field fluorescence- and/or TIRF illumination system, C1, C1Si spectral confocal or LiveScan Sweptfield confocal system offer a wide variety of imaging capabilities. CLEM can be used with the C1 to reduce photobleaching and provides greater flexibility in time-lapse studies. Nikon,s TE2000 series microscopes include noise-terminator technology to improve signal-to-noise rations. A number of specialized fluorescence objectives are available (Plan Fluor, Super Fluor, Plan Apochromat, Plan Apochromat VC).
