Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes

König, Karsten; So, Peter T. C.; Mantulin, William W.; Gratton, Enrico

doi:10.1364/ol.22.000135

Cited by 249 publications

(180 citation statements)

References 6 publications

(6 reference statements)

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“…2F with Fig. 1A and E), supporting the proposal that mitochondria are the initial, and possibly the major, source of reactive oxygen in response to damaging light 17,23,31,32 . This increase was not due to endogenous autofluorescence, as unstained embryos do not exhibit this increase after irradiation (Fig.…”

Section: Peroxide Is Produced In Lscm-imaged Embryossupporting

confidence: 79%

“…Our results also show that the arrest is not due to alterations in the culture medium, as unimaged embryos in the same drop remain viable. Cellular damage caused by light has been shown in other cell types for various wavelengths, ranging from 295 to 500 nm (one photon) 22,23 or 730 nm to 800 nm (two photons) 17 . Therefore, we sought to determine a possible endogenous source of this phototoxicity.…”

Section: Author Manuscript Author Manuscriptmentioning

confidence: 99%

“…However, it is still unclear whether TPLSM is a suitable technique for long-term imaging studies of living preparations because exposure to the very high intensity illumination of TPLSM may be damaging to living cells. It has been demonstrated that TPLSM mean excitation doses at levels above 6 mW and 150 fs at wavelengths of ≤800 nm can compromise tissue culture cell viability 17 . Furthermore, concerns have been raised that the longer wavelengths used for TPLSM may cause localized heating of the specimen 18,19 .…”

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Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability

et al. 1999

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A major challenge for fluorescence imaging of living mammalian cells is maintaining viability following prolonged exposure to excitation illumination. We have monitored the dynamics of mitochondrial distribution in hamster embryos at frequent intervals over 24 h using two-photon microscopy (1,047 nm) while maintaining blastocyst, and even fetal, developmental competence. In contrast, confocal imaging for only 8 h inhibits development, even without fluorophore excitation. Photo-induced production of H 2 O 2 may account, in part, for this inhibition. Thus, twophoton microscopy, but not confocal microscopy, has permitted long-term fluorescence observations of the dynamics of three-dimensional cytoarchitecture in highly photosensitive specimens such as mammalian embryos.Keywords two-photon microscopy; laser scanning confocal microscopy; live cell fluorescence imaging; embryo; mitochondrial dynamics; mammal; hamster The detection of specific cellular components by imaging techniques such as wide-field epifluorescence or laser scanning confocal microscopy (LSCM) requires exposure to high intensity light that can cause cellular damage 1 . Consequently, the quantity or quality of images that can be collected is limited or, even worse, the reliability of the images may be compromised. This is a particular problem when imaging events that occur over periods of time ranging from hours to days, such as embryonic development. For this reason, much of our current understanding of subcellular morphological changes during mammalian embryonic development is based on images of fixed or static specimens at different developmental stages [2][3][4][5][6] . Thus, it can be difficult to interpret dynamic processes accurately, because the continuity of events must be inferred. The establishment of long-term fluorescence imaging methods that maintain the viability of live specimens is critical for advancing our understanding of cell biology and embryonic development in areas such as ion dynamics 7 , cytoplasmic reorganization, compaction and blastocoel formation, embryonic development in exotic species (where specimens are heterogeneous and difficult * Corresponding author (jsquirre@students.wisc.edu). to obtain), use of fluorescent tags for the preselection of embryos for subsequent embryo transfer 8 , and studies of protein expression in living cells using green fluorescent protein 9 . HHS Public AccessEmbryos of some mammals, particular hamsters, are very sensitive to culture conditions 10 . Furthermore, studies suggest that mammalian oocytes and embryos are adversely affected by exposure to visible light [11][12][13] . Because of this sensitivity, mammalian embryos are ideal to test live-cell imaging techniques. In addition, there are obvious morphological changes associated with differentiation, namely compaction and blastocoel formation, which can be used to assess viability. The embryo must undergo cell division during and after imaging as well as maintain a level of developmental competence that allows it to initiate dif...

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Section: Peroxide Is Produced In Lscm-imaged Embryossupporting

confidence: 79%

Section: Author Manuscript Author Manuscriptmentioning

confidence: 99%

mentioning

confidence: 99%

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Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability

et al. 1999

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“…These lasers can supply high irradiance in the range of GW cm À2 for transient durations as short as 10 À9 to 10 À15 seconds within the focal volume. Even so, during an ultrashort pulse duration, high peak irradiance delivered may still damage the irradiated cells by eliciting a variety of undesired biological responses [4][5][6][7][8][9][10]. However, the mentioned harmful cellular effects are restricted to the irradiated tissue and its immediate surroundings and do not have long term effects.…”

Section: Biophotonicsmentioning

confidence: 99%

Estimating the risk of squamous cell cancer induction in skin following nonlinear optical imaging

Thomas

Nadiarnykh

Voskuilen

et al. 2013

Journal of Biophotonics

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High power femto‐second (fs) laser pulses used for in‐vivo nonlinear optical (NLO) imaging can form cyclobutane pyrimidine dimers (CPD) in DNA, which may lead to carcinogenesis via subsequent mutations. Since UV radiation from routine sun exposure is the primary source of CPD lesions, we evaluated the risk of CPD‐related squamous cell carcinoma (SCC) in human skin due to NLO imaging relative to that from sun exposure. We developed a unique cancer risk model expanding previously published estimation of risk from exposure to continuous wave (CW) laser. This new model showed that the increase in CPD‐related SCC in skin from NLO imaging is negligible above that due to regular sun exposure. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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“…Damage threshold-The issue of photodamage by high-intensity illumination has been extensively studied in the context of two-photon fluorescence microscopy (101,102). The damage mechanism associated with high instantaneous intensity light illumination has been conjectured to be due to the destructive intracellular plasma formation or the destructive singlet oxygen formation (or both) and indirect DNA damage.…”

Section: Light Fluence Considerationsmentioning

confidence: 99%

Molecular Contrast Optical Coherence Tomography: A Review

Yang¹

2004

Photochem Photobiol

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This article reviews the current state of research on the use of molecular contrast agents in optical coherence tomography (OCT) imaging techniques. After a brief discussion of the basic principle of OCT and the importance of incorporating molecular contrast agent usage into this imaging modality, we shall present an overview of the different molecular contrast OCT (MCOCT) methods that have been developed thus far. We will then discuss several important practical issues that define the possible range of contrast agent choice, the design criteria for engineered molecular contrast agent and the implementability of a given MCOCT method for clinical or biological applications. We will conclude by outlining a few areas of pursuit that deserve a greater degree of research and development.

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Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes

Cited by 249 publications

References 6 publications

Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability

Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability

Estimating the risk of squamous cell cancer induction in skin following nonlinear optical imaging

Molecular Contrast Optical Coherence Tomography: A Review

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