Quantum Yield Determination Based on Photon Number Measurement, Protocols for Firefly Bioluminescence Reactions

  • Kazuki NiwaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1461)


Quantum yield (QY), which is defined as the probability of photon production by a single bio/chemiluminescence reaction, is an important factor to characterize luminescence light intensity emitted diffusively from the reaction solution mixture. Here, methods to measure number of photons to determine QY according to the techniques of national radiometry standards are described. As an example, experiments using firefly bioluminescence reactions are introduced.

Key words

Firefly luciferase Quantum yield Bioluminescence Luminometer National radiometry standard 



K.N. would like to thank Y. Ichino for helpful discussions about absolute calibrations and Y. Ohmiya for his discussions about bioluminescence reactions. This work was supported by a Grant-in-Aid from the Japan Society for the Promotion of Science.


  1. 1.
    Seliger HH, McElroy WD (1959) Quantum yield in the oxidation of firefly luciferin. Biochem Biophys Res Commun 1:21–24CrossRefGoogle Scholar
  2. 2.
    Ando Y, Niwa K, Yamada N, Enomoto T, Irie T, Kubota H, Ohmiya Y, Akiyama H (2007) Firefly bioluminescence quantum yield and color change by pH-sensitive green emission. Nat Photon 2:44–47CrossRefGoogle Scholar
  3. 3.
    Niwa K, Ichino Y, Ohmiya Y (2010) Quantum yields measurements of firefly bioluminescence reactions using a commercial lumnometer. Chem Lett 39:291–293CrossRefGoogle Scholar
  4. 4.
    Niwa K, Ichino Y, Kumata S, Nakajima Y, Hiraishi Y, Kato D, Viviani RV, Ohmiya Y (2010) Quantum yields and kinetics of the firefly bioluminescence reaction of beetle luciferases. Photochem Photobiol 86:1046–1049CrossRefPubMedGoogle Scholar
  5. 5.
    Seliger HH, McElroy WD, White EH, Field GF (1961) Stereospecificity and firefly bioluminescence, a comparison of natural and synthetic luciferins. Proc Natl Acad Sci U S A 47:1129–1134CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Zong Y, Miller CC, Lykke KR, Ohno Y (2004) Measurement of total radiant flux of UV LEDs. Proc CIE CIE x026:107–110Google Scholar
  7. 7.
    Wood KV (1995) The chemical mechanism and evolutionary development of beetle bioluminescence. Photochem Photobiol 62:662–673CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Quantum Optical Measurement Group, National Metrology Institute of JapanNational Institute of Advanced Industrial Science and TechnologyTsukubaJapan

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