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Qualification of a fluorescence spectrometer for measuring true fluorescence spectra
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10.1063/1.2715952
/content/aip/journal/rsi/78/3/10.1063/1.2715952
http://aip.metastore.ingenta.com/content/aip/journal/rsi/78/3/10.1063/1.2715952

Figures

Image of FIG. 1.
FIG. 1.

Schematics of the instrumental setups used for measuring signals and determining spectral correction.

Image of FIG. 2.
FIG. 2.

The relative intensity (primary axis) of the excitation beam at the sample position as a function of wavelength, measured using a calibrated detector (corrected) and the channel (reference). The relative -channel correction factor (secondary axis) as a function of excitation (ex) wavelength ( correction). The axis is normalized to one at for both corrected and reference curves.

Image of FIG. 3.
FIG. 3.

The relative intensity (primary axis) of the calibrated source as a function of wavelength, including the calibrated values (CS) and those measured by the channel ( channel). The relative -channel correction factor (secondary axis) as a function of emission (em) wavelength ( correction). The axis is normalized to one at for both CS and -channel curves.

Image of FIG. 4.
FIG. 4.

The relative -channel correction factor (primary axis) as a function of em wavelength, as calculated using the data measured with CS-based and CD-based methods. The percent difference between and (secondary axis) as a function of em wavelength. The axis is normalized to one at for both and .

Image of FIG. 5.
FIG. 5.

Comparison of contour plotted EEMs for tryptophan in aqueous solution, where the fluorescence intensity is (a) the measured uncorrected signal, maxima at and , (b) the signal corrected for the excitation intensity or flux at the sample position, maximum at , and (c) the instrument-independent signal, corrected for both the instrument’s excitation flux and its spectral responsivity, maximum at .

Image of FIG. 6.
FIG. 6.

The absolute -channel correction factor (primary axis) as a function of em wavelength at an ex wavelength of , as calculated using the data measured with CS-based and CD-based methods. The percent difference between and (secondary axis) as a function of em wavelength.

Image of FIG. 7.
FIG. 7.

The absolute -channel correction factor (primary axis) as a function of ex wavelength at an em wavelength of , as calculated using the data measured with CS-based and CD-based methods. The percent difference between and (secondary axis) as a function of ex wavelength.

Image of FIG. 8.
FIG. 8.

A plot of the -channel intensity at as a function of the percent transmittance of the calibrated filter used to attenuate the intensity reaching the detector. The line is a linear least squares fit of the points in the linear intensity range of the PMT.

Image of FIG. 9.
FIG. 9.

A plot of the bias in the measured em wavelengths relative to the actual values for Hg, Kr, and Ne atomic lines. The curve is a nonlinear least squares fit of the points to a fourth order polynomial. Lines are used to connect groups of points where sinusoidal behavior is observed.

Image of FIG. 10.
FIG. 10.

The and factors of the excitation beam and the detection system, respectively, as a function of wavelength.

Tables

Generic image for table
Table I.

Atomic line positions used for accuracy validation.

Generic image for table
Table II.

Estimated uncertainties in the relative correction factors and .

Generic image for table
Table III.

Estimated uncertainties in the absolute correction factor .

Generic image for table
Table IV.

Wavelengths used for accuracy validation and observed bias in nm.

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/content/aip/journal/rsi/78/3/10.1063/1.2715952
2007-03-26
2014-04-16
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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Qualification of a fluorescence spectrometer for measuring true fluorescence spectra
http://aip.metastore.ingenta.com/content/aip/journal/rsi/78/3/10.1063/1.2715952
10.1063/1.2715952
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