The data provided by Spectra Viewer can be used only for preliminary estimation of a relative reporter's suitability for your instrumentation and experimental system. The actual performance depends on many additional parameters, such as expression levels, protein turnover rates, type and relative power of the excitation sources, etc. The real suitability of a particular fluorescent protein for your task should be tested experimentally.
Non-normalized spectra
Non-normalized view allows one to compare the excitation and emission efficiencies of different FPs between each other within the selected wavelength range. Here the amplitude of each excitation peak is equal to the maximal molar extinction coefficient of the corresponding FP, and the total area of each emission spectrum is equal to the fluorescence quantum yield of the corresponding FP.
Wide-field excitation
Ex: Relative efficiency of excitation by light of the selected wavelength range. The value is calculated as the ratio of the area under the excitation curve limited by selected wavelengths to the total area of excitation spectrum of FusionRed (the reporter with the largest area of excitation spectrum among currently available Evrogen FPs).
Em: Quantum yield of corresponding fluorescent protein within the selected wavelength range. The value is calculated for each particular FP as a portion of the area under the emission curve, limited by selected wavelengths.
Σ: Product of Ex and Em, X 10000. The value reflects the total brightness of the signal, collected in the corresponding channel. The value is shown in red in the occurrence of an inappropriate combination of excitation and emission bandpasses in the same channel.
Laser excitation
Ex: Efficiency of excitation by chosen laser line. The value corresponds to the molar extinction coefficient at the selected wavelength.
Em: Quantum yield of a corresponding fluorescent protein within the selected wavelength range. The value is calculated for each particular FP as a portion of the area under the emission curve, limited by selected wavelengths.
Σ: Product of Ex and Em, divided by 1000. The value corresponds to the calculated brightness of the signal, collected in the corresponding channel. The value is shown in red in the occurrence of an inappropriate combination of excitation and emission wavelengths in the same channel.
Normalized spectra
Normalized view shows the relative efficiencies of excitation and emission within the selected wavelength range as a portion of maximal efficiency for each reporter. Here the amplitude of each excitation and emission spectra is normalized to 100%.
Wide-field excitation
Ex: Relative efficiency of excitation by light of the selected wavelength range. The value is calculated for each particular FP as a portion of the area under the excitation curve, limited by selected wavelengths.
Em: Portion of emission light gathered within the selected wavelength range. The value is calculated for each particular FP as a portion of the area under the emission curve, limited by selected wavelengths.
Σ: Product of Ex and Em, X 100. The value corresponds to the percentage of the signal, collected in the corresponding channel. The value is shown in red in the occurrence of an inappropriate combination of excitation and emission bandpasses in the same channel.
Laser excitation
Ex: Relative efficiency of excitation by chosen laser line.
Em: Portion of emission light gathered within the selected wavelength range. The value is calculated for each particular FP as a portion of the area under the emission curve, limited by selected wavelengths.
Σ: Product of Ex and Em, X 100. The value corresponds to the percentage of the signal, collected in the corresponding channel. The value is shown in red in the occurrence of an inappropriate combination of excitation and emission wavelengths in the same channel.
