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Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes
Abstract
The influence of the pulse length, tau , of ultrashort laser pulses at 780 and 920 nm on cell vitality and cellular reproduction has been studied. A total of 2400 nonlabeled cells were exposed to a highly focused scanning beam from a mode-locked 80-MHz Ti:sapphire laser with 60-micros pixel dwell time. For the same pulse energy, destructive effects were more pronounced for shorter pulses. The damage behavior was found to follow approximately a P(2)/tau dependence (P , mean power), indicating that cell destruction is likely based on a two-photon excitation process rather than a one- or a three-photon event. Therefore, femtosecond as well as picosecond pulses provide approximately the same relative optical window for safe two-photon fluorescence microscopy.
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Cited by
Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission
Klar, T. A., Jakobs, S., Dyba, M., Egner, A., & Hell, S. W. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.97.15.8206 · Cited by 1,869
Multiphoton microscopy in life sciences
König, K. (2000) · Journal of Microscopy · doi:10.1046/j.1365-2818.2000.00738.x · Cited by 1,265
Self-citationPrinciples of two-photon excitation microscopy and its applications to neuroscience
Svoboda, K., & Yasuda, R. (2006) · Neuron · doi:10.1016/j.neuron.2006.05.019 · Cited by 1,027
Phototoxicity in live fluorescence microscopy, and how to avoid it
Icha, J., Weber, M., Waters, J. C., & Norden, C. (2017) · BioEssays · doi:10.1002/bies.201700003 · Cited by 622
Highly Nonlinear Photodamage in Two-Photon Fluorescence Microscopy
Hopt, A., & Neher, E. (2001) · Biophysical Journal · doi:10.1016/s0006-3495(01)76173-5 · Cited by 442
Metadata
- Authors
- König, K., Becker, T. W., Fischer, P., Riemann, I., & Halbhuber, K.-J.
- Venue
- Optics Letters
- Publisher
- Optica Publishing Group
- Published
- 1999-01-15
- Volume
- 24
- Issue
- 2
- Pages
- 113
- ISSN
- 0146-9592 (ISSN-L)
- 1539-4794
- OpenAlex
- W2040815295
- Cited by
- 259
- References
- 0
- FWCI
- 10.86
- Type
- journal article
- Language
- en
- Open access
- closed
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- König, K., Becker, T. W., Fischer, P., Riemann, I., & Halbhuber, K.-J. (1999). Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes. Optics Letters. https://doi.org/10.1364/ol.24.000113
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