The DOI matches its Crossref record; 26 of the 27 references it cites were found in a registry or an index.
Highly Nonlinear Photodamage in Two-Photon Fluorescence Microscopy
Open accessAbstract
Two-photon fluorescence excitation is being increasingly used in laser scan microscopy due to very low photodamage induced by this technique under normal operation. However, excitation intensity has to be kept low, because nonlinear photodamage sets in when laser power is increased above a certain threshold. We studied this kind of damage in bovine adrenal chromaffin cells, using two different indicators of damage: changes in resting [Ca(2+)] level and the degranulation reaction. In agreement with previous studies, we found that, for both criteria, damage is proportional to the integral (over space and time) of light intensity raised to a power approximately 2.5. Thus, widening the laser pulse shape at constant average intensity both in time and in focal volume is beneficial for avoiding this kind of damage. Both measures, of course, reduce the two-photon fluorescence excitation. However, loss of signal can be compensated by increasing excitation power, such that, at constant damaging potential, signals may be even larger with long pulses and large focal volumes, because the exponent of the power law of damage is higher (mu approximately 2.5) than that of the two-photon signal (mu approximately 2).
Overview
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“This uncaging regime produced little or no photodamage29, as shown by the stability of the amplitudes of the 2pEPSCs over 1 min (Fig. 1h).”
From Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons (2001) · cites this paper as [29]
Citations
Cites
Two-photon laser scanning fluorescence microscopy
Denk, W., Strickler, J. H., & Webb, W. W. (1990) · Science · doi:10.1126/science.2321027 · Cited by 9,488
Handbook of fluorescent probes and research chemicals
Haugland, R. P. (1996) · Medical Entomology and Zoology · Cited by 2,793
Xu, C., & Webb, W. W. (1996) · Journal of the Optical Society of America B · doi:10.1364/josab.13.000481 · Cited by 2,198
Ultrashort Laser Pulse Phenomena
Diels, J., & Rudolph, W. (2006) · Elsevier eBooks · doi:10.1016/b978-0-12-215493-5.x5000-9 · Cited by 853
Theory and Practice of Scanning Optical Microscopy
Dainty, J. C. (1985) · Optica Acta International Journal of Optics · doi:10.1080/713821694 · Cited by 773
Lynch, D. K. (1987) · Journal of the Optical Society of America A · doi:10.1364/josaa.4.000558 · Cited by 756
Photon Upmanship: Why Multiphoton Imaging Is More than a Gimmick
Denk, W. T., & Svoboda, K. (1997) · Neuron · doi:10.1016/s0896-6273(00)81237-4 · Cited by 679
Breakdown threshold and plasma formation in femtosecond laser–solid interaction
von der Linde, D., & Schüler, H. (1996) · Journal of the Optical Society of America B · doi:10.1364/josab.13.000216 · Cited by 308
Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage
Koester, H. J., Baur, D., Uhl, R., & Hell, S. W. (1999) · Biophysical Journal · doi:10.1016/s0006-3495(99)77063-3 · Cited by 265
König, K., Becker, T., Fischer, P., Riemann, I., & Halbhuber, K. J. (1999) · Optics Letters · doi:10.1364/ol.24.000113 · Cited by 259
Book Review: Ultrashort Laser Pulse Phenomena
Miller, S. A. (1997) · Optical Engineering · doi:10.1117/1.601465 · Cited by 259
Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes
König, K., So, P. T. C., Mantulin, W. W., & Gratton, E. (1997) · Optics Letters · doi:10.1364/ol.22.000135 · Cited by 255
Williams, R. M., Piston, D. W., & Webb, W. W. (1994) · The FASEB Journal · doi:10.1096/fasebj.8.11.8070629 · Cited by 236
Heating by absorption in the focus of an objective lens
Schönle, A., & Hell, S. W. (1998) · Optics Letters · doi:10.1364/ol.23.000325 · Cited by 192
Cell damage by near-IR microbeams
König, K., Liang, H., Berns, M. W., & Tromberg, B. J. (1995) · Nature · doi:10.1038/377020a0 · Cited by 170
Cell damage in near-infrared multimode optical traps as a result of multiphoton absorption
König, K., Liang, H., Berns, M. W., & Tromberg, B. J. (1996) · Optics Letters · doi:10.1364/ol.21.001090 · Cited by 158
Two-photon near- and far-field fluorescence microscopy with continuous-wave excitation
Hell, S. W., Booth, M. J., Wilms, S., Schnetter, C., Kirsch, A. K., Arndt‐Jovin, D. J., & Jovin, T. M. (1998) · Optics Letters · doi:10.1364/ol.23.001238 · Cited by 83
Fast scanning and efficient photodetection in a simple two-photon microscope
Tan, Y. P., Llano, I., Hopt, A., Würriehausen, F., & Neher, E. (1999) · Journal of Neuroscience Methods · doi:10.1016/s0165-0270(99)00103-x · Cited by 67
Self-citationSmith, C. L. (1999) · Journal of Neuroscience · doi:10.1523/jneurosci.19-02-00589.1999 · Cited by 64
Quantitative analysis of exocytosis directly visualized in living chromaffin cells
Terakawa, S., Fan, J., Kumakura, K., & Ohara‐Imaizumi, M. (1991) · Neuroscience Letters · doi:10.1016/0304-3940(91)90163-n · Cited by 56
König, K., Simon, U., & Halbhuber, K. J. (1996) · PubMed · Cited by 50
Booth, & Hell, S. W. (1998) · Journal of Microscopy · doi:10.1046/j.1365-2818.1998.00375.x · Cited by 46
Picosecond pulsed two‐photon imaging with repetition rates of 200 and 400 MHz
Bewersdorf, & Hell, S. W. (1998) · Journal of Microscopy · doi:10.1046/j.1365-2818.1998.00379.x · Cited by 45
Femtosecond pulse width control in microscopy by two‐photon absorption autocorrelation
Brakenhoff, G. J., Mueller, M., & Squier, J. A. (1995) · Journal of Microscopy · doi:10.1111/j.1365-2818.1995.tb03638.x · Cited by 25
Light and living matter : a guide to the study of photobiology.
Clayton, R. K. (1970) · Cited by 21
Image formation in three-photon fluorescence microscopy
Sheppard, C. J. R. (1996) · Bioimaging · doi:10.1002/1361-6374(199609)4:3 · Cited by 20
Research works printed but not linked: 2
- Clayton, 1977, Vol. 1 and 2
- Ridsdale, 1993, The viability of cultured cells under two-photon laser scanning microscopy (abstr.), Biophys. J., 64, A109
Cited by
Sorted by citations
Deep tissue two-photon microscopy
Helmchen, F., & Denk, W. T. (2005) · Nature Methods · doi:10.1038/nmeth818 · Cited by 4,288
Nonlinear magic: multiphoton microscopy in the biosciences
Zipfel, W. R., Williams, R. M., & Webb, W. W. (2003) · Nature Biotechnology · doi:10.1038/nbt899 · Cited by 3,911
Matsuzaki, M., Ellis-Davies, G. C. R., Nemoto, T., Miyashita, Y., Iino, M., & Kasai, H. (2001) · Nature Neuroscience · doi:10.1038/nn736 · Cited by 1,563
Mechanisms of femtosecond laser nanosurgery of cells and tissues
Vogel, A., Noack, J., Hüttman, G., & Paltauf, G. (2005) · Applied Physics B · doi:10.1007/s00340-005-2036-6 · Cited by 1,445
Principles 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
Metadata
- Authors
- Hopt, A., & Neher, E.
- Affiliations
- Max Planck Institute for Biophysical Chemistry (DE)
- Max Planck Society (DE)
- Corresponding author
- Neher, Erwin
- ORCID
- Neher, Erwin
- Venue
- Biophysical Journal
- Publisher
- Elsevier BV
- Published
- 2001-04
- Volume
- 80
- Issue
- 4
- Pages
- 2029-2036
- ISSN
- 0006-3495 (ISSN-L)
- PMC
- PMC1301392
- OpenAlex
- W2089975180
- Cited by
- 442
- References
- 27
- Citations by year
- 2026: 15 · 2025: 15 · 2024: 15 · 2023: 16 · 2022: 23 · 2021: 21 · 2020: 16 · 2019: 14 · 2018: 14 · 2017: 18 · 2016: 16 · 2015: 18 · 2014: 24 · 2013: 18 · 2012: 13
- FWCI
- 9.13
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biophysical Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Materials Science › Electronic, Optical and Magnetic Materials › Nonlinear Optical Materials Studies
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Laser-Matter Interactions and Applications
- Keywords
- two-photon fluorescence microscopy, photodamage, laser pulse shape, focal volume, excitation intensity, adrenal chromaffin cells, calcium imaging, degranulation
- MeSH terms
- Adrenal Glands, Animals, Calcium, Cattle, Lasers, Light, Microscopy, Fluorescence, Models, Theoretical, Time Factors, Models, Statistical, Photons, Chromaffin Cells
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
