| Popis: |
The first isolated and most commonly used fluorescent protein is the green fluorescent protein (GFP). However, fluorescent proteins that cover the whole visible spectrum have been available and used as indispensable tools in a large spectrum of bioscience research. Fluorescent microscopes illuminate samples with the light of one colour, but detect only the light emitted by the fluorophores in the sample. Microscopes have been constantly developing, and one of the major advancements was the use of light emitting diodes (LED) as the source of light. The advantages of LEDs are uniform illumination, less dissipated heat and an extremely long life time. However, LEDs are limited to certain fixed wavelengths unlike the previously used arch lamps that emit white light. Therefore, the aim of this study was to determine the compatibility of ten commonly used fluorescent proteins (mTagBFP2, mTFP1, mEGFP, mCitrine, tdTomato, mTagRFP, mCherry, mKate, mPlum, and E2Crimson) with the EVOS Floid Imagining Station fluorescence microscope which uses blue, red and green light emitting diodes (LEDs) as a light source. We found out that, although the EVOS Floid Imagining Station fluorescence microscope has only three LEDs emitting specific wavelengths of the spectrum, it can detect a wide range of fluorescent proteins. Out of ten tested proteins only the mTagRFP was not detected. Furthermore, it is possible to distinguish five proteins in the same sample, one blue (mTagBFP2), one turquoise (mTFP1), one orange (mKate), one of three green (mEGFP, mCitrine or tdTomato) and one of three red (mCherry, mPlum or E2 Crimson) fluorescent proteins. |
