High-Speed Fluorescence Imaging and Intensity Profiling of Femtosecond-Induced Calcium Transients
We have demonstrated a combined imaging system, where the physiology of biological specimens can be imaged and profiled at 10–20 frames per second whilst undergoing femtosecond laser irradiation. Individual GH3 cells labeled with the calcium fluorophore Fluo-3 were stimulated using a counter-propagating focused femtosecond beam with respect to the imaging system. As a result of the stimulation, calcium waves can be generated in COS cells, and laser-induced calcium oscillations are initiated in the GH3 cells. Single-photon fluorescence images and intensity profiles of the targeted specimens are sampled in real-time using a modified PerkinElmer UltraView LCI microscope.
- J. White and E. H. Stelzer, “Photobleaching GFP reveals protein dynamics inside live cells,” Trends in Cell Biology, vol. 9, no. 2, pp. 61–65, 1999.
- U. K. Tirlapur and K. König, “Technical advance: near-infrared femtosecond laser pulses as a novel non-invasive means for dye-permeation and 3D imaging of localised dye-coupling in the Arabidopsis root meristem,” The Plant Journal, vol. 20, no. 3, pp. 363–370, 1999.
- U. K. Tirlapur and K. König, “Cell biology: targeted transfection by femtosecond laser,” Nature, vol. 418, no. 6895, pp. 290–291, 2002.
- H. Oehring, I. Riemann, P. Fischer, K.-J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning, vol. 22, no. 4, pp. 263–270, 2000.
- U. K. Tirlapur and K. König, “Femtosecond near-infrared laser pulses as a versatile non-invasive tool for intra-tissue nanoprocessing in plants without compromising viability,” The Plant Journal, vol. 31, no. 3, pp. 365–374, 2002.
- K. König, I. Riemann, P. Fischer, and K.-J. Halbhuber, “Intracellular nanosurgery with near infrared femtosecond laser pulses,” Cellular and Molecular Biology, vol. 45, no. 2, pp. 195–201, 1999.
- K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, and E. Gratton, “Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes,” Optics Letters, vol. 22, no. 2, pp. 135–136, 1997.
- K. König, T. W. Becker, P. Fischer, I. Riemann, and K.-J. Halbhuber, “Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes,” Optics Letters, vol. 24, no. 2, pp. 113–115, 1999.
- M. W. Berns, J. Aist, J. Edwards et al., “Laser microsurgery in cell and developmental biology,” Science, vol. 213, no. 4507, pp. 505–513, 1981.
- N. Shen, M. Colvin, F. Genin et al., “Using femtosecond laser subcellular surgery to study cell biology,” Biophysical Journal, vol. 86, supplment S, pp. 520A–520A, 2004.
- B. C. Stuart, M. D. Feit, S. Herman, A. M. Rubenchik, B. W. Shore, and M. D. Perry, “Optical ablation by high-power short-pulse lasers,” Journal of the Optical Society of America B: Optical Physics, vol. 13, no. 2, pp. 459–468, 1996.
- W. Watanabe, N. Arakawa, S. Matsunaga et al., “Femtosecond laser disruption of subcellular organelles in a living cell,” Optics Express, vol. 12, no. 18, pp. 4203–4213, 2004.
- S. K. Mohanty, M. Sharma, and P. K. Gupta, “Laser-assisted microinjection into targeted animal cells,” Biotechnology Letters, vol. 25, no. 11, pp. 895–899, 2003.
- W. Tao, J. Wilkinson, E. J. Stanbridge, and M. W. Berns, “Direct gene transfer into human cultured cells facilitated by laser micropuncture of the cell membrane,” Proceedings of the National Academy of Sciences of the United States of America, vol. 84, no. 12, pp. 4180–4184, 1987.
- V. Venugopalan, A. Guerra III, K. Nahen, and A. Vogel, “Role of laser-induced plasma formation in pulsed cellular microsurgery and micromanipulation,” Physical Review Letters, vol. 88, no. 7, p. 078103, 2002.
- K. König, “Laser tweezers are sources of two-photon excitation,” Cellular and Molecular Biology, vol. 44, no. 5, pp. 721–733, 1998.
- B. Agate, C. T. A. Brown, W. Sibbett, and K. Dholakia, “Femtosecond optical tweezers for in-situ control of two-photon fluorescence,” Optics Express, vol. 12, no. 13, pp. 3011–3017, 2004.
- N. I. Smith, K. Fujita, T. Kaneko et al., “Generation of calcium waves in living cells by pulsed-laser-induced photodisruption,” Applied Physics Letters, vol. 79, no. 8, pp. 1208–1210, 2001.
- H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, “ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage,” Biophysical Journal, vol. 77, no. 4, pp. 2226–2236, 1999.
- A. Ichihara, T. Tanaami, K. Isozaki et al., “High-speed confocal fluorescence microscopy using a Nipkow scanner with microlenses for 3D-imaging of single fluorescent molecule in real time,” Bioimages, vol. 4, no. 1, pp. 52–62, 1996.
- P. E. Hänninen and S. W. Hell, “Femtosecond pulse broadening in the focal region of a two-photon fluorescence microscope,” Bioimaging, vol. 2, no. 3, pp. 117–121, 1994.
- M. Müller, J. Squier, and G. J. Brakenhoff, “Measurement of femtosecond pulses in the focal point of a high-numerical-aperture lens by two-photon absorption,” Optics Letters, vol. 20, no. 9, pp. 1038–1040, 1995.
- D. Bird and M. Gu, “Two-photon fluorescence endoscopy with a micro-optic scanning head,” Optics Letters, vol. 28, no. 17, pp. 1552–1554, 2003.
- A. H. Tashjian Jr., Y. Yasumura, L. Levine, G. H. Sato, and M. L. Parker, “Establishment of clonal strains of rat pituitary tumor cells that secrete growth hormone,” Endocrinology, vol. 82, no. 2, pp. 342–352, 1968.
Copyright © 2006 Daniel Day et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.