Fluorophores
A fluorophore is a fluorescent chemical that can re-emit light upon excitation. Fluorophores can be used in conjugation with protein or oligonucleotides to generate fluorescent markers for detecting the expression of proteins and nucleic acid. Fluorescent labels functionally accept light energy of a given wavelength (e.g., from a laser) and re-emit energy at longer wavelength. These two processes are known as excitation and emission.
Different types of fluorophores are used in flow cytometry based on the instrument setting (see lasers and detection channels in Figure 1), the feature of fluorophores, and markers needed in designated panels such as four, six, or ten color panels.
Figure 1 Brand names of Flow Cytometer and its laser and channel setting.
Figure 2 Caprico Biotechnologies' 10 Color Panel, and the Spectrum of Most Used Fluorophores.
Dyes such as FITC, PE, APC and PerCP have become the standard for many years. Tandem dyes, comprising a small fluorophore covalently coupled to a larger fluorophore, provide maximum utilization of specific equipment, increased detection channels for each laser, and generate more information for valuable, sometimes very limited, samples.
Commonly used fluorophores for surface or intracellular epitope detection are described in Figure 2.
There are many fluorophores that have potential applications in flow cytometry. A new generation of small molecule fluorophores such as Alexa Fluor, iFluor, and mFluor are rapidly replacing many traditional fluorophores, providing users with greater light stability and brighter fluorescence. CBI offers these high-performance fluorescent dyes: iFluor™ and mFluor™ series.
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