Journal of Fluorescence

, Volume 23, Issue 3, pp 605–610 | Cite as

Decomposition of Complex Fluorescence Spectra Containing Components with Close Emission Maxima Positions and Similar Quantum Yields. Application to Fluorescence Spectra of Proteins

  • Aleksandar Savić
  • Roland Kardos
  • Miklós Nyitrai
  • Ksenija Radotić


Despite of widely application of multivariate analysis in chemometrics, problem of resolving closely positioned components in the fluorescence spectra remained unsolved, thus limiting the usage of fluorescence spectroscopy in analytical purpose. In this paper we have described a novel procedure, adapted especially for the analysis of complex fluorescence spectra with multiple, closely positioned components’ maxima. The method was first tested on the simulated spectra and then applied on the spectra of proteins whose fluorophores have similar properties of both the excitation and the emission spectra. In this paper, simple but efficient modification of the method was applied. Instead of analyzing full size emission matrix (12 spectra), 9 spectra wide windows were analyzed, and 4 factors (greatest possible number of factors with physical meaning both for actin and simulated spectra) were extracted in each pass. Obtained factor scores were grouped by using the K-means algorithm. Groups of factor scores obtained from K-means algorithm were passed through the one more factor analysis (FA) in order to find one factor that represents each group. Our approach provides resolution of extremely closed spectral components, which is a vital data for protein conformation analysis based on fluorescence spectroscopy.


Fluorescence spectra Proteins Fixed size window factor analysis Clustering 



Grant 173017 from the Ministry of Education and Science of the Republic of Serbia supported this study. We thank to Mr Dragosav Mutavdžić for critical reading of this paper. This study was supported by the Hungarian Science Foundation (OTKA grant K77840 to MN) and also by the ‘Science, Please! Research Team on Innovation’ (SROP-4.2.2/08/1/2008-0011).


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Aleksandar Savić
    • 1
  • Roland Kardos
    • 2
  • Miklós Nyitrai
    • 2
    • 3
  • Ksenija Radotić
    • 1
  1. 1.Institute for multidisciplinary researchUniversity of BelgradeBelgradeSerbia
  2. 2.Department of BiophysicsUniversity of Pecs, Medical SchoolPecsHungary
  3. 3.Szentágothai Research CenterPécsHungary

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