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The excitation and emission spectra of fluorescent molecules are commonly observed to overlap. The difference in wavelength or energy between the excitation and emitted fluorescent photons is called the Stokes shift. In practice, the Stokes shift is the difference between the excitation and emission maxima (Fig. 11-3). Depending on the particular fluorescent molecule, the shift can range from just a few to several hundred nanometers. The Stokes shift for fluorescein is 20 nm, while that for porphyrins is over 200 nm. Dyes exhibiting a large Stokes shift are advantageous in fluorescence microscopy, because the bands of excitation and fluorescence wavelengths are easier to isolate using interference filters. Another important criterion for dye selection is the molar extinction coefficient, which describes the potential of a fluorochrome to absorb photon quanta, and is given in units of absorbance (optical density) at a reference wavelength (usually the absorption maximum) under specified conditions. The quantum efficiency (QE) of fluorescence emission is the fraction of absorbed photon quanta that is re-emitted by a fluorochrome as fluorescent photons. QE varies greatly between different fluorochromes and for a single fluorochrome under different conditions. For soluble fluorescein dye at alkaline pH, the quantum efficiency can be as high as 0.9 an extremely vb.net upc-a reader VB . NET UPC-A Barcode Reader SDK - Decode & Scan UPC-A ...
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