OSA's Digital Library

Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 6 — Jun. 1, 2012
  • pp: 1266–1273

Light-induced Ca2+ transients observed in widefield epi-fluorescence microscopy of excitable cells

Alison McDonald, John Harris, Debbi MacMillan, John Dempster, and Gail McConnell  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 6, pp. 1266-1273 (2012)

View Full Text Article

Enhanced HTML    Acrobat PDF (1168 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



We have investigated the possibility that variations in the level of intracellular Ca2+ in excitable cells might be induced as an artifact of the incoherent illumination that is being used to monitor transient responses. In order to avoid the fluctuations in power of an arc lamp source, a microscope using a light emitting diode that was calibrated accurately at low power levels, was constructed to provide good control over the dose of light applied to the biological specimen. We report here that higher powers of illumination increased the probability of occurrence of Ca2+ transients even in the sub-mW range normally used to measure such transients in epi-fluorescence work, suggesting that caution should be exercised when designing experiments and interpreting data.

© 2012 OSA

OCIS Codes
(170.0110) Medical optics and biotechnology : Imaging systems
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(170.5380) Medical optics and biotechnology : Physiology

ToC Category:

Original Manuscript: March 2, 2012
Revised Manuscript: April 25, 2012
Manuscript Accepted: April 25, 2012
Published: May 4, 2012

Alison McDonald, John Harris, Debbi MacMillan, John Dempster, and Gail McConnell, "Light-induced Ca2+ transients observed in widefield epi-fluorescence microscopy of excitable cells," Biomed. Opt. Express 3, 1266-1273 (2012)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. N. C. Shaner, P. A. Steinbach, and R. Y. Tsien, “A guide to choosing fluorescent proteins,” Nat. Methods2(12), 905–909 (2005). [CrossRef] [PubMed]
  2. J. Zhang, R. E. Campbell, A. Y. Ting, and R. Y. Tsien, “Creating new fluorescent probes for cell biology,” Nat. Rev. Mol. Cell Biol.3(12), 906–918 (2002). [CrossRef] [PubMed]
  3. R. W. Cole and J. N. Turner, “Light-emitting diodes are better illumination sources for biological microscopy than conventional sources,” Microsc. Microanal.14(03), 243–250 (2008). [CrossRef] [PubMed]
  4. D. F. Albeanu, E. Soucy, T. F. Sato, M. Meister, and V. N. Murthy, “LED arrays as cost effective and efficient light sources for widefield microscopy,” PLoS ONE3(5), e2146 (2008). [CrossRef] [PubMed]
  5. V. Ntziachristos, “Going deeper than microscopy: the optical imaging frontier in biology,” Nat. Methods7(8), 603–614 (2010). [CrossRef] [PubMed]
  6. J. White and E. Stelzer, “Photobleaching GFP reveals protein dynamics inside live cells,” Trends Cell Biol.9(2), 61–65 (1999). [CrossRef] [PubMed]
  7. J. Lippincott-Schwartz, E. Snapp, and A. Kenworthy, “Studying protein dynamics in living cells,” Nat. Rev. Mol. Cell Biol.2(6), 444–456 (2001). [CrossRef] [PubMed]
  8. S. Iwanaga, N. I. Smith, K. Fujita, and S. Kawata, “Slow Ca(2+) wave stimulation using low repetition rate femtosecond pulsed irradiation,” Opt. Express14(2), 717–725 (2006). [CrossRef] [PubMed]
  9. N. I. Smith, S. Iwanaga, T. Beppu, K. Fujita, O. Nakamura, and S. Kawata, “Photostimulation of two types of Ca2+ waves in rat pheochromocytoma PC12 cells by ultrashort pulsed near-infrared laser irradiation,” Laser Phys. Lett.3(3), 154–161 (2006). [CrossRef]
  10. M. M. Knight, S. R. Roberts, D. A. Lee, and D. L. Bader, “Live cell imaging using confocal microscopy induces intracellular calcium transients and cell death,” Am. J. Physiol. Cell Physiol.284(4), C1083–C1089 (2003). [PubMed]
  11. J. G. McCarron and T. C. Muir, “Mitochondrial regulation of the cytosolic Ca2+ concentration and the InsP3-sensitive Ca2+ store in guinea-pig colonic smooth muscle,” J. Physiol.516(1), 149–161 (1999). [CrossRef] [PubMed]
  12. S. Inoue, “Foundations of confocal scanned imaging in light microscopy,” in Handbook of Biological Confocal Microscopy, J.B Pawley, ed. (Plenum, 1995), pp. 1–19.
  13. J. Dempster, D. Wokosin, K. McCloskey, J. Girkin, and A. Gurney, “WinFluor-an integrated system for the simultaneous recording of cell fluorescence images and electrophysiological signals on a single computer system,” Br. J. Pharmacol.137(Suppl. S), 146P (2002).
  14. R. Lavi, A. Shainberg, H. Friedmann, V. Shneyvays, O. Rickover, M. Eichler, D. Kaplan, and R. Lubart, “Low energy visible light induces reactive oxygen species generation and stimulates an increase of intracellular calcium concentration in cardiac cells,” J. Biol. Chem.278(42), 40917–40922 (2003). [CrossRef] [PubMed]
  15. W. D. McElroy and B. Glass, A Symposium on Light and Life (John Hopkins Press, Baltimore, 1961).
  16. A. Schönle and S. W. Hell, “Heating by absorption in the focus of an objective lens,” Opt. Lett.23(5), 325–327 (1998). [CrossRef] [PubMed]
  17. N. I. Kiskin, R. Chillingworth, J. A. McCray, D. Piston, and D. Ogden, “The efficiency of two-photon photolysis of a “caged” fluorophore, o-1-(2-nitrophenyl)ethylpyranine, in relation to photodamage of synaptic terminals,” Eur. Biophys. J.30(8), 588–604 (2002). [CrossRef] [PubMed]
  18. A. C. Giese, Blepharisma, the Biology of a Light-Sensitive Protozoan (Stanford University Press, Stanford, Calif, 1973).
  19. Y. J. Suzuki and G. D. Ford, “Redox regulation of signal transduction in cardiac and smooth muscle,” J. Mol. Cell. Cardiol.31(2), 345–353 (1999). [CrossRef] [PubMed]
  20. R. Lubart, H. Friedmann, and R. Lavie, “Photobiostimulation as a function of different wavelengths,” Laser Therapy12(1), 38–41 (2000). [CrossRef]
  21. R. Dixit and R. Cyr, “Cell damage and reactive oxygen species production induced by fluorescence microscopy: effect on mitosis and guidelines for non-invasive fluorescence microscopy,” Plant J.36(2), 280–290 (2003). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.


Fig. 1 Fig. 2 Fig. 3
Fig. 4 Fig. 5

Supplementary Material

» Media 1: MPG (3162 KB)     

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited