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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 4, Iss. 4 — Apr. 1, 2009

Real-time imaging of autofluorescence NAD(P)H in single human neutrophils

Leiting Pan, Xinzheng Zhang, Kun Song, Baiquan Tang, Wei Cai, Xian Wu, Romano A. Rupp, and Jingjun Xu  »View Author Affiliations

Applied Optics, Vol. 48, Issue 6, pp. 1042-1046 (2009)

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Real-time detection of NAD(P)H is particularly important for understanding physiological activities of neutrophils. We scrutinize the performance of weak light detection systems with electron multiplying CCDs (EMCCDs) with regard to the feasibility of valid investigations by autofluorescence NAD(P)H in single human neutrophils. The low-noise amplification facility of EMCCDs is indeed just adequate to permit detection at an irradiation level where neither quenching nor phototoxic effects occur. For demonstration, a neutrophil respiratory burst was triggered and observed in real time. Our low-intensity detection system fulfills all requirements for real-time investigations at high spatiotemporal resolution in the field of neutrophil physiology and pathology.

© 2009 Optical Society of America

OCIS Codes
(110.2970) Imaging systems : Image detection systems
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(260.2510) Physical optics : Fluorescence

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: August 26, 2008
Revised Manuscript: November 20, 2008
Manuscript Accepted: January 27, 2009
Published: February 12, 2009

Virtual Issues
Vol. 4, Iss. 4 Virtual Journal for Biomedical Optics

Leiting Pan, Xinzheng Zhang, Kun Song, Baiquan Tang, Wei Cai, Xian Wu, Romano A. Rupp, and Jingjun Xu, "Real-time imaging of autofluorescence NAD(P)H in single human neutrophils," Appl. Opt. 48, 1042-1046 (2009)

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  1. W. Ying, “NAD+/NADH and NADP+/NADPH in cellular functions and cell death: regulation and biological consequences,” Antioxid. Redox. Signal. 10, 179-206 (2008). [CrossRef]
  2. W. Kuhn, T. Müller, R. Winkel, S. Danielczik, A. Gerstner, R. Häcker, C. Mattern, and H. Przuntek, “Parenteral application of NADH in Parkinson's disease: clinical improvement partially due to stimulation of endogenous levodopa biosynthesis,” J. Neural Transm. 103, 1187-1193 (1996). [CrossRef] [PubMed]
  3. A. Mayevsky and G. G. Rogatsky, “Mitochondrial function in vivo evaluated by NADH fluorescence: from animal models to human studies,” Am. J. Physiol. Cell. Physiol. 292, C615-C640 (2007). [CrossRef]
  4. J. R. Zhang, K. Vrecko, K. Nadlinger, D. Storga, G. D. Birkmayer, and G. Reibnegger, “The reduced coenzyme nicotinamide adenine dinucleotide (NADH) repairs DNA damage of PC12 cells induced by doxorubicin,” J. Tumor Marker Oncol. 13, 5-17 (1998).
  5. B. W. Pogue, J. D. Pitts, M. A. Mycek, R. D. Sloboda, C. M. Wilmot, J. F. Brandsema, and J. A. O'Hara, “In vivo NADH fluorescence monitoring as an assay for cellular damage in photodynamic therapy,” Photochem. Photobiol. 74, 817-824 (2001). [CrossRef]
  6. M. Hashimoto, Y. Takeda, T. Sato, H. Kawahara, O. Nagano, and M. Hirakawa, “Dynamic changes of NADH fluorescence images and NADH content during spreading depression in the cerebral cortex of gerbils,” Brain Res. 872, 294-300 (2000). [CrossRef] [PubMed]
  7. M. Dellinger, M. Geze, R. Santus, E. Kohen, C. Kohen, J. G. Hirschberg, and M. Monti, “Imaging of cells by autofluorescence: a new tool in the probing of biopharmaceutical effects at the intracellular level,” Biotechnol. Appl. Biochem. 28, 25 (1998). [PubMed]
  8. T. E. Decoursey, V. V. Cherny, W. Zhou, and L. L. Thomas, “Simultaneous activation of NADPH oxidase-related proton and electron currents in human neutrophils,” Proc. Natl. Acad. Sci. U.S.A. 97, 6885-6889 (2000). [CrossRef] [PubMed]
  9. T. E. Wientjes and A. W. Segal, “NADPH oxidase and the respiratory burst,” Semin. Cell. Biol. 6, 357-365(1995). [CrossRef] [PubMed]
  10. T. Mair, C. Warnke, and S. C. Müller, “Spatio-temporal dynamics in glycolysis,” Faraday Discuss. 120, 249-259(2002). [CrossRef] [PubMed]
  11. D. W. Piston and S. M. Knobel, “Real-time analysis of glucose metabolism by microscopy,” Trends Endocrinol. Metab. 10, 413-417 (1999). [CrossRef] [PubMed]
  12. C. G. Coates, D. J. Denvir, E. Conroy, N. G. McHale, K. D. Thornbury, and M. A. Hollywood, “Back-illuminated electron multiplying technology: The world's most sensitive CCD for ultra low-light microscopy,” http://www.emccd.com/emccd_in_use/publications_and_scientific_papers.
  13. C. G. Coates, D. J. Denvir, N. G. McHale, K. D. Thornbury, and M. A. Hollywood, “Ultra-sensitivity, speed and resolution: Optimizing low-light microscopy with the back-illuminated electron multiplying CCD,” http://www.emccd.com/emccd_in_use/publications_and_scientific_papers.
  14. C. Nathan, “Neutrophils and immunity: challenges and opportunities,” Nat. Rev. Immunol. 6, 173-182 (2006). [CrossRef] [PubMed]

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