Kit Spectral Fluorescence Standards: Principle and Properties

The Kit Spectral Fluorescence Standards consists of

- five spectral fluorescence standards covering the spectral region of 300 to 730 nm
- certified normalized corrected emission spectra
- solvent ethanol
- software LINKCORR
- data sheet with procedure

The five standards are carefully designed to match a set of requirements regarding spectral properties.

The straightforward working principle is illustrated in Figure 3.

- Addition of 10 ml of ethanol to each of the fluorescent standards yields working solutions with
  absorbances of ca. 0.04 at the maximum of the dye's lowest energy absorption band.
- These solutions are measured with the instrument to be calibrated.
- Data evaluation is performed with included software LINKCORR


LINCORR calculates the quotients Q^F00x(λ_em) of the certified emission spectra Ic(λ_em) (Fig. 3, bottom, solid lines) and the uncorrected emission spectra Iu(λ_em) (Fig. 3, bottom, dashed lines) for each dye. Weighted combination of Q^F00x(λ_em), shown in the middle part of Fig. 3, by LINKCORR yields an overall emission correction curve (Fig. 3, top). This curve equals the inverse relative spectral responsivity of the instrument 1/s(λ_em). Corrected instrument-independent and comparable data are then obtained upon multiplication of measured spectra with this correction curve (1/s(λ_em)).

Working principle of the Kit Spectral Fluorescence Standards. Division of the certified corrected spectra of standard dyes (Ic(λem), bottom, solid lines) by the recorded uncorrected spectra (Iu(λem), bottom, dashed lines) of these dyes leads to individual quotients QF00x(λem), see middle part. Weighted combination of QF00x(λem) by LINKCORR yields an overall emission correction curve for the chosen measurement conditions of the instrument to be calibrated that equals the instrument´s inverse relative spectral responsivity 1/s(λem). Corrected and thus comparable spectra are obtained upon subsequent multiplication of measured spectra with the correction curve.

Certified Fluorescence Standard
Instrument Effects on Fluorescence Signals
Determination of the Spectral Responsivity
Tracability, Certification, and Proviniency Testing
References

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