Aria 8.0 Podręcznik Użytkownika Pobierz pdf (Strona 49)

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Chapter 2: Theory of Operation 49

Fluorescent Signals

When cells or particles stained with fluorochrome-conjugated antibodies or other

dyes pass through a laser beam, the dyes can absorb photons (energy) and be

promoted to an excited electronic state. In returning to their ground state, the

dyes release energy, most of which is emitted as light. This light emission is

known as fluorescence.

Fluorescence is always a longer wavelength (lower-energy photon) than the

excitation wavelength. The difference between the excitation wavelength and the

emission wavelength is known as the Stokes shift. Some fluorescent compounds

such as PerCP exhibit a large Stokes shift, absorbing blue light (488 nm) and

emitting red light (675 nm), while other fluorochromes such as FITC have a

smaller Stokes shift, absorbing blue light and emitting green light (530 nm).

The emission spectra for some commonly used fluorochromes are shown in

Figure 2-5. See Table 8-7 on page 258 for the corresponding detectors.

Figure 2-5 Emission spectra of commonly used fluorochromes

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