Abstract
With the Calibration Kit Spectral Fluorescence Standards BAM-F001–BAM-F005, we developed a simple tool for the characterization of the relative spectral responsivity and the long-term stability of the emission channel of fluorescence instruments under routine measurement conditions thereby providing the basis for an improved comparability of fluorescence measurements and eventually standardization. This first set of traceable fluorescence standards, which links fluorescence measurements to the spectral radiance scale in the spectral range of 300–770 nm and has been optimized for spectrofluorometers, can be employed for different measurement geometries and can be adapted to different fluorescence techniques with proper consideration of the underlying measurement principles.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Notes
NIST: National Institute of Standards and Technology, USA; NRC: National Research Council, Canada, PTB: Physikalisch-Technische Bundesanstalt, Germany; BAM: Federal Institute for Materials Research and Testing, Germany.
BAM-F001–BAM-F005: dyes A–E in earlier publications.
Due to radiometric convention, s(λ) always implies s λ(λ).
In the case of ratio mode-type instruments that record the quotient of the signal of the emission and the reference channel, both detectors need to be operated within their linear range.
References
Lakowicz JR (1999) Principles of fluorescence spectroscopy, 2nd edn. Kluwer Academic/Plenum, New York
Lakowicz JR (ed) (1992–2004) Topics in fluorescence spectroscopy, series vol 1–8. Plenum, New York
Wolfbeis OS (series ed) (2001–2004) Springer series on fluorescence, methods and applications, vol 1–3. Springer, Berlin
Schulman SG (ed) (1985–1993) Molecular luminescence spectroscopy parts 1–3. Wiley Interscience, New York
Mason WT (1999) Fluorescent and luminescent probes for biological activity, 2nd edn. Academic, San Diego
Resch-Genger U, Hoffmann K, Nietfeld W, Engel A, Neukammer J, Nitschke R, Ebert B, Macdonald R (2005) How to improve quality assurance in fluorometry: fluorescence-inherent sources of error and suited fluorescence standards. J Fluoresc 15(3):337–362 and references therein
Monte C, Resch-Genger U, Pfeifer D, Taubert RD, Hollandt J (2005) Linking fluorescences measurements to radiometric units, Metrologia (in press)
Resch-Genger U, Pfeifer D, Monte C, Pilz W, Hoffmann A, Spieles M, Rurack K, Hollandt J, Taubert D, Schönenberger B, Nording P (2005) Traceability in fluorometry: part II. Spectral fluorescence standards. J Fluoresc 15(3):315–336
Certificate of the Federal Institute for Materials Research and Testing (BAM) (2006) Certified Reference Materials BAM-F001, BAM-F002, BAM-F003, BAM-F004, and BAM-F005, Calibration Kit Spectral Fluorescence Standards, for the determination of the relative spectral responsivity of fluorescence instruments, January 10, 2006
Hollandt J, Taubert DR, Seidel J, Resch-Genger U, Gugg-Helminger A, Pfeifer D Monte C (2005) Traceability in fluorometry: part I physical standards. J Fluoresc 15(3):301–313
Eastman JW (1966) Standardization of fluorescence spectra and the calibration of spectrofluorimeters. Appl Opt 5(7):1125–1132
Miller JN (1981) Standards in fluorescence spectrometry. Ultraviolett Spectrometry Group, London
Velapoldi RA, Tonnesen HH (2004) Corrected emission spectra and quantum yields for a series of fluorescent compounds in the visible spectral region. J Fluoresc 14(4):465–472
Hofstraat JW, Latuhihin MJ (1994) Correction of fluorescence-spectra. Appl Spectrom 48(4):436–447
Hoffmann K, Monte C, Pfeifer D, Resch-Genger U (2005) Standards in fluorescence spectroscopy: simple tools for the characterization of fluorescence instruments. GIT Lab J 5(3):2–4
Velapoldi RA, Mielenz KD (1980) A fluorescence standard reference material: quinine sulfate dihydrate. NBS Special Publication, PB 80132046, Springfield, VA, pp 260-264
Gardecki JA, Maroncelli M (1998) Set of secondary emission standards for calibration of the spectral responsivity in emission spectroscopy. Appl Spectrom 52(9):1179–1189
Hoffmann K, Resch-Genger U, Nitschke R (2005) Simple Tool for the Standardization of Confocal Spectral Imaging Systems. GIT Imaging Microsc (3):18–19
Bremser W, de Rose P, Duewer DS, Ebert B, Hoffmann A, Hollandt J, Macdonald R, Monte C, Pfeifer D, Resch-Genger U, Taubert DR, Zwinkels J (manuscript in preparation)
Pfeifer D, Hoffmann A, Monte C, Spieles M, Bremser W, Resch-Genger U (manuscript in preparation
Acknowledgments
Financial support from the German Ministry of Economics and Labor (BMWA) is gratefully acknowledged (grants BMWA VI A2–18 and VI A2–17/03). We express our gratitude to Dr J. Hollandt and Dr R. D. Taubert from PTB for the calibration of the spectral radiance transfer standards and provision of the spectral radiance of the tungsten ribbon lamp displayed in Fig. 2, to Dr A. Höpe from PTB for the calibration of the diffuse reflectance standard, and to Dr B. Schönenberger and Dipl.-Biol. P. Nording from Sigma-Aldrich GmbH (former Fluka Production GmbH) for the supply of the kit dyes and the solvent. We are also very grateful for the analytical characterization of the kit dyes by our BAM colleagues Dr U. Erhardt, Dr P. Lehnink-Habrink, Dipl.-Chem. O. Hahnebeck, and Mrs R. Schmieder (chromatography), Dr M. Koch and Mrs E. Tzscheutschler (water content, Karl Fischer titration), and Dr F. Malz (quantitative NMR) as well as to Dr W. Bremser for help with the determination of the uncertainty budget for the corrected emission spectra of the kit components.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pfeifer, D., Hoffmann, K., Hoffmann, A. et al. The Calibration Kit Spectral Fluorescence Standards—A Simple and Certified Tool for the Standardization of the Spectral Characteristics of Fluorescence Instruments. J Fluoresc 16, 581–587 (2006). https://doi.org/10.1007/s10895-006-0086-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10895-006-0086-8