Fluorescence-intensity distribution analysis and its application in biomolecular detection technology

DOI Web Site 被引用文献2件
  • Peet Kask
    EVOTEC BioSystems AG, Schnackenburgallee 114, D-22525 Hamburg, Germany; and Institute of Experimental Biology, Instituudi tee 11, EE3051 Harku, Estonia
  • Kaupo Palo
    EVOTEC BioSystems AG, Schnackenburgallee 114, D-22525 Hamburg, Germany; and Institute of Experimental Biology, Instituudi tee 11, EE3051 Harku, Estonia
  • Dirk Ullmann
    EVOTEC BioSystems AG, Schnackenburgallee 114, D-22525 Hamburg, Germany; and Institute of Experimental Biology, Instituudi tee 11, EE3051 Harku, Estonia
  • Karsten Gall
    EVOTEC BioSystems AG, Schnackenburgallee 114, D-22525 Hamburg, Germany; and Institute of Experimental Biology, Instituudi tee 11, EE3051 Harku, Estonia

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<jats:p>A methodology, fluorescence-intensity distribution analysis, has been developed for confocal microscopy studies in which the fluorescence intensity of a sample with a heterogeneous brightness profile is monitored. An adjustable formula, modeling the spatial brightness distribution, and the technique of generating functions for calculation of theoretical photon count number distributions serve as the two cornerstones of the methodology. The method permits the simultaneous determination of concentrations and specific brightness values of a number of individual fluorescent species in solution. Accordingly, we present an extremely sensitive tool to monitor the interaction of fluorescently labeled molecules or other microparticles with their respective biological counterparts that should find a wide application in life sciences, medicine, and drug discovery. Its potential is demonstrated by studying the hybridization of 5′-(6-carboxytetramethylrhodamine)-labeled and nonlabeled complementary oligonucleotides and the subsequent cleavage of the DNA hybrids by restriction enzymes.</jats:p>

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