Linking fluorescence spectroscopy to the scale of spectral sensitivity: the BAM reference fluorometer

Christian Monte; Walter Pilz; Ute Resch-Genger

doi:10.1117/12.628912

18 August 2005 Linking fluorescence spectroscopy to the scale of spectral sensitivity: the BAM reference fluorometer

Christian Monte, Walter Pilz, Ute Resch-Genger

Proceedings Volume 5880, Optical Diagnostics; 588019 (2005) https://doi.org/10.1117/12.628912
Event: Optics and Photonics 2005, 2005, San Diego, California, United States

Abstract

Providing fluorescence and fluorescence excitation spectra traceable to the scale of spectral sensitivity (responsivity) and spectral radiance at minimized uncertainty is currently limited by two factors: The uncertainty of the available transfer standards and the uncertainty of the measurement process itself. Here the requirements on a reference fluorometer enabling measurements at minimized uncertainty, its design, the simulation and the realization are presented. The fluorometer is designed with minimized chromatic and geometrical aberrations. To realize an efficient reduction of stray light and subtractive dispersion a double monochromator design was necessary. The basic element is a so-called U-type Czerny-Turner single monochromator featuring off-axis parabolas and an entrance and exit slit virtually at the same place. Thereby spherical aberration, coma and astigmatism are effectively minimized. The here employed special double monochromator design further cancels the remaining aberrations of the single monochromator. The design of the whole spectrometer was optimized with a ray tracing program. To minimize uncertainties due to the transfer standards, the reference fluorometer is exclusively traceable to the spectral sensitivity (responsivity) scale. This enables the use of transfer standards with much smaller uncertainty. Here trap detectors are employed of common design but specially calibrated for a divergent light bundle. Based on this instrument with its achromatic design and precisely known numerical apertures the determination of absolute fluorescence spectra will be addressed.

Citation Download Citation

Christian Monte, Walter Pilz, and Ute Resch-Genger "Linking fluorescence spectroscopy to the scale of spectral sensitivity: the BAM reference fluorometer", Proc. SPIE 5880, Optical Diagnostics, 588019 (18 August 2005); https://doi.org/10.1117/12.628912

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