Skip to main content
U.S. flag

An official website of the United States government

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.

State-of-the-Art Comparability of Corrected Emission Spectra-Part II: Peer-toPeer Assessment of Calibration Performance Using Spectral Fluorescence Standards

Published

Author(s)

Paul C. DeRose, Joanne C. Zwinkels, Bernd Ebert

Abstract

The tremendous growth of fluorescence applications in the life and material sciences has proceeded largely without sufficient concern for the reliability and uncertainties related to the characterization and performance validation of fluorescence instruments and measurements. To assess the state-of-the-art comparability and trueness of corrected emission spectra accomplishable with the aid of commercially available spectral fluorescence standards, in part II of this study, twelve field laboratories were randomly selected from academia and industry based upon their fluorescence measuring equipment. These laboratories performed a reference material (RM)-based fluorometer calibration following a standard operation procedure provided with the fluorescence standards and used this emission correction curve for the determination of the corrected emission spectra of three test dyes X, QY, and Y. This revealed an average accuracy of the correctedemission spectra of the test dyes of minimum 6.8 % for common commercial spectrofluorometers, routine measurement conditions, and the data evaluation software LINKCORR developed and provided by BAM. These results only slightly exceed the relative standard uncertainties of 4.2 % for physical standard (PTS)-based spectral corrections and 2.4 % for RM-based spectral corrections demonstrated in part I of this study by an international interlaboratory comparison of four expert laboratories.
Citation
Analytical Chemistry
Volume
84

Keywords

biosystems and health, calibration, fluorescence, instrument qualification, method validation, reference materials, spectral correction

Citation

DeRose, P. , Zwinkels, J. and Ebert, B. (2012), State-of-the-Art Comparability of Corrected Emission Spectra-Part II: Peer-toPeer Assessment of Calibration Performance Using Spectral Fluorescence Standards, Analytical Chemistry, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=907887 (Accessed April 24, 2024)
Created February 28, 2012, Updated February 19, 2017