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Fluorescent dyes (or fluorochromes) are commonly used as detection reagents in various applications such as cellular imaging and flow cytometry. Fluorochromes absorb light energy of a specific wavelength and re-emit it at a longer wavelength. The wavelengths at which the fluorochrome absorbs and emits light are known as the fluorochrome's excitation and emission spectra, respectively (or just fluorescence spectra).Â
Instruments requiring the use of fluorescent dyes, such as fluorescence microscopes and flow cytomeres, are equipped with lasers producing light at a particular wavelength so they can excite fluorochromes capable of absorbing light at that specific wavelength. The light emitted by the fluorochrome is then filtered so that each sensor will detect fluorescence only within the filter's range. This fluorescence is the read-out signal provided by the instrument.
Many different fluorochromes have been developed, each one with a particular emission and excitation fluorescence spectra. You can find commercially available fluorescent dyes that absorb and emit light at specific wavelengths across the whole visible spectrum, including the infrared region. Fluorochromes differ in the intensity at which they emit light. Therefore, the fluorochrome brightness will depend on its ability to absorb light and the efficiency at which the absorbed light is converted into emitted light. Â
Selecting the right fluorochrome may seem difficult if you do not have adequate tools. The task becomes even more complicated when designing a multiplex panel that requires two or more fluorochromes. In this case, you must consider the spectral overlap between fluorochromes and other factors to minimize potential problems such as fluorescence spillover.
Our comprehensive fluorochrome chart will guide you through each of the steps involved in selecting a fluorochrome. Featuring the 30 most popular labels, our chart will allow you to quickly choose what fluorochromes are the most suitable for your next multiplex experiment.