Journal of Fluorescence

, Volume 22, Issue 1, pp 381–389 | Cite as

Fluorescence Detection by Intensity Change Based Sensors: A Theoretical Model

  • Javier Galbán
  • Arantzazu Delgado-Camón
  • Vicente L. Cebolla
  • Susana de Marcos
  • Víctor Polo
  • Elena Mateos
Original Paper


According to Fluorescence Detection by Intensity Changes (FDIC) the fluorescence intensity of many fluorophores depends on the non-covalent (specific and/or non-specific) interactions these fluorophores would be able to establish with the solvent and, more interestingly, with other surrounding molecules. This latter effect is the basis of FDIC for analytical purposes. In this paper, a preliminary study of FDIC applications using a fluorophore supported in a solid medium (sensor film) is presented. First, a mathematical model relating the analyte concentration with the immobilized fluorophore fluorescence is deduced. The model includes all the different mechanisms explaining this relationship: index of refraction or dielectric constant modification, scattering coefficient alteration and sensor film volume increase. Then, the very first experimental results are presented, using different fluorophores and solid supports. The best results were obtained using polyacrylamide (PAA) polymers and coralyne as the fluorophore. This sensor film is applied for albumin and polyethylenglycol determination and the results are compared with those obtained using coralyne in solution. Albumin quenches the coralyne fluorescence in both cases (solution and film), while PEG quenches coralyne fluorescence in films but increases it in solution. These results suggest that the outstanding fluorescence change mechanism is sensor films is the film volume increases, which is different than those observed in solution.


FDIC Non-covalent interactions Sensors Polyethylenglycol Albumin 



Authors thank the Spanish Ministerio de Ciencia e Innovacion (MICIN) of Spain (projects CTQ 2008-06751-C02-01/BQU and CTQ 2008–00959). A.D-C. thanks to the MICIN for a grant.

Supplementary material

10895_2011_970_MOESM1_ESM.doc (189 kb)
ESM 1 (DOC 189 kb)


  1. 1.
    Lakowicz JR (1999) Principles of fluorescence spectroscopy, Chapter 6, 2nd edn. Kluwer Academic/Plenum Press, New York, pp 185–237Google Scholar
  2. 2.
    Valeur B (2202) Molecular fluorescence: principles and applications, Wiley-VCH, Chapter 7, pp. 200–225.Google Scholar
  3. 3.
    Cossıo FP, Arrieta A, Cebolla VL, Membrado L, Domingo MP, Henrion P, Vela J (2000) Enhancement of fluorescence in thin-layer chromatography induced by the interaction between n-Alkanes and an organic Cation. Anal Chem 72:1759–1766PubMedCrossRefGoogle Scholar
  4. 4.
    Cossıo FP, Arrieta A, Cebolla VL, Membrado L, Vela J, Garriga R, Domingo MP (2000) Berberine Cation: a fluorescent chemosensor for Alkanes and other low-polarity compounds. An explanation of this phenomenon. Org Lett 2:2311–2313PubMedCrossRefGoogle Scholar
  5. 5.
    Brown MB, Miller JN, Seare NJ (1995) An investigation of the use of Nile Red as a long-wavelength fluorescent probe for the study of α1-acid glycoprotein-drug interactions. J Pharm Biomed Anal 13:1011–1017PubMedCrossRefGoogle Scholar
  6. 6.
    Li W, Lu Z (1998) The fluorescent re action between Berberine and DNA and the fluorometry of DNA. Microchem J 60:84–88CrossRefGoogle Scholar
  7. 7.
    Gong GQ, Zong ZX, Song YM (1999) Spectrofluorometric determination of DNA and RNA with berberine, Spectrochim. Acta 55A:1903–1907Google Scholar
  8. 8.
    Cser A, Nagy K, Biczok L (2002) Fluorescence lifetime of Nile Red as a probe for the hydrogen bonding strength with its microenvironment. Chem Phys Lett 360:473–478CrossRefGoogle Scholar
  9. 9.
    Gálvez E, Matt M, Cebolla VL, Fernándes F, Membrado L, Cossıo FP, Garriga R, Vela J, Guermouche H (2006) General contribution of nonspecific interactions to fluorescence intensity. Anal Chem 78:3699–3705PubMedCrossRefGoogle Scholar
  10. 10.
    Galbán J, Mateos E, Cebolla V, Domínguez A, Delgado-Camón A, de Marcos S, Sanz-Vicente I, Sanz V (2009) The environmental effect on the fluorescence intensity in solution. An analytical model. Analyst 134:2286–2292PubMedCrossRefGoogle Scholar
  11. 11.
    Sanz V, de Marcos S, Galbán J (2007) A reagentless optical biosensor based on the intrinsic absorption properties of peroxidase. Biosens Bioelectron 22:956–964PubMedCrossRefGoogle Scholar
  12. 12.
    Cass AEG (1990) Biosensors. A practical approach (Practical Approach Series). Oxford University Press, OxfordGoogle Scholar
  13. 13.
    Cooper J, Cass A (2004) Biosensors. A practical approach, 2nd edn. Oxford, Oxford University PressGoogle Scholar
  14. 14.
    Becke AD (1988) Density-functional exchange-energy approximation with correct asymptotic-behaviour. Phys Rev A 38:3098–32100PubMedCrossRefGoogle Scholar
  15. 15.
    Becke AD (1993) Density-functional thermochemistry. 3. The role of exact exchange. J Chem Phys 98:5648–5652CrossRefGoogle Scholar
  16. 16.
    Lee CT, Yang WT, Parr RG (1988) Development of the Colle-Salvetti correlation-energy formula into a functional of the electron-density. Phys Rev B 37:785–789CrossRefGoogle Scholar
  17. 17.
    Ditchfield R, Hehre WJ, Pople JA (1970) Molecular orbital theory of bond separation. J Chem Phys 52:13–14CrossRefGoogle Scholar
  18. 18.
    Galbán J, Delgado-Camón A, Sanz V, Sanz-Vicente I, de Marcos S (2008) A theoretical approach for designing fluorescent biosensors: The optical model. Anal Chim Acta 615:148–157PubMedCrossRefGoogle Scholar
  19. 19.
    see supplementary materialGoogle Scholar
  20. 20.
    Yankov D (2004) Diffusion of glucose and maltose in polyacrylamide gel. Enz Microb Technol 34:603–610CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Javier Galbán
    • 1
  • Arantzazu Delgado-Camón
    • 2
  • Vicente L. Cebolla
    • 2
  • Susana de Marcos
    • 1
  • Víctor Polo
    • 3
  • Elena Mateos
    • 1
  1. 1.Analytical Biosensors Group, INA (Institute of Nanoscience of Aragón), Analytical Chemistry Department, Faculty of ScienceUniversity of ZaragozaZaragozaSpain
  2. 2.Group of Chemical Technology for Separation and DetectionInstitute of Carboquimica. CSICZaragozaSpain
  3. 3.Physical-Chemistry Department, Faculty of ScienceUniversity of ZaragozaZaragozaSpain

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