Short and long-term phototoxicity in cells expressing genetic reporters under nanosecond laser exposure

Gottschalk, Sven; Estrada, Héctor; Degtyaruk, Oleksiy; Rebling, Johannes; Klymenko, Olena; Rosemann, Michael; Razansky, Daniel

doi:10.1016/j.biomaterials.2015.07.051

Cited by 8 publications

(11 citation statements)

References 38 publications

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“…It was previously shown that pulsed light radiation may cause photobleaching of fluorescent probes and other photodegradation mechanisms for energy levels below safety standards [270]. Even though these effects are generally considered detrimental, they can be exploited for enhancing the resolution and contrast in optoacoustic microscopy by considering the signal difference before and after photodegradation [34].…”

Section: Dynamic Contrast Enhancement Approachesmentioning

confidence: 99%

“…The light pulses employed in optoacoustic imaging induce photobleaching effects that considerably differ from those known from fluorescence imaging using single- or multi-photon microscopes [270, 280]. It has also been long recognized that pulsed laser radiation has different biological effects from those observed under continuous wave exposure [281].…”

Section: Dynamic Contrast Enhancement Approachesmentioning

confidence: 99%

“…Furthermore, FPs are known to induce phototoxicity under certain excitation conditions [285]. While most of these effects are yet unexplored for realistic in vivo imaging conditions, optoacoustic signal degradation associated to photobleaching effects in FPs has already been observed under different imaging settings [270, 280] (Fig. 12b).…”

Section: Dynamic Contrast Enhancement Approachesmentioning

confidence: 99%

“…(c) Loss of fluorescence signal (shown in %) due to photobleaching of mCherry-expressing cells under different illumination conditions (average intensity and fluence) within ANSI exposure limits for 10 second exposure with 10000 pulses. Adapted with permission from [270]. © 2015 Elsevier.…”

Section: Figmentioning

confidence: 99%

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Advanced optoacoustic methods for multiscale imaging of in vivo dynamics

et al. 2017

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Visualization of dynamic functional and molecular events in an unperturbed in vivo environment is essential for understanding the complex biology of living organisms and of disease state and progression. To this end, optoacoustic (photoacoustic) sensing and imaging have demonstrated the exclusive capacity to maintain excellent optical contrast and high resolution in deep-tissue observations, far beyond the penetration limits of modern microscopy. Yet, the time domain is paramount for the observation and study of complex biological interactions that may be invisible in single snapshots of living systems. This review focuses on the recent advances in optoacoustic imaging assisted by smart molecular labeling and dynamic contrast enhancement approaches that enable new types of multiscale dynamic observations not attainable with other bio-imaging modalities. A wealth of investigated new research topics and clinical applications is further discussed, including imaging of large-scale brain activity patterns, volumetric visualization of moving organs and contrast agent kinetics, molecular imaging using targeted and genetically expressed labels, as well as three-dimensional handheld diagnostics of human subjects.

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Section: Dynamic Contrast Enhancement Approachesmentioning

confidence: 99%

Section: Dynamic Contrast Enhancement Approachesmentioning

confidence: 99%

Section: Dynamic Contrast Enhancement Approachesmentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

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Advanced optoacoustic methods for multiscale imaging of in vivo dynamics

et al. 2017

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“…However, nowadays, the interaction of visible light with living cells is still of great interest for a wide variety of purposes [5][6][7][8]. Among a plethora of cell features used for investigating cell damages, the morphology and volume are gaining great significance.…”

Section: Introductionmentioning

confidence: 99%

Investigating fibroblast cells under “safe” and “injurious” blue‐light exposure by holographic microscopy

Calabuig

Mugnano

Miccio

et al. 2016

Journal of Biophotonics

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The exposure to visible light has been shown to exert various biological effects, such as erythema and retinal degeneration. However, the phototoxicity mechanisms in living cells are still not well understood. Here we report a study on the temporal evolution of cell morphology and volume during blue light exposure. Blue laser irradiation is switched during the operation of a digital holography (DH) microscope between what we call here "safe" and "injurious" exposure (SE & IE). The results reveal a behaviour that is typical of necrotic cells, with early swelling and successive leakage of the intracellular liquids when the laser is set in the "injurious" operation. In the phototoxicity investigation reported here the light dose modulation is performed through the very same laser light source adopted for monitoring the cell's behaviour by digital holographic microscope. We believe the approach may open the route to a deep investigation of light-cell interactions, with information about death pathways and threshold conditions between healthy and damaged cells when subjected to light-exposure. 3D Morphology and quantitative phase information from late stage of necrosis cell death.

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High-Throughput Platform for Optoacoustic Probing of Genetically Encoded Calcium Ion Indicators

et al. 2019

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SummaryFunctional optoacoustic (OA) imaging assisted with genetically encoded calcium ion indicators (GECIs) holds promise for imaging large-scale neuronal activity at depths and spatiotemporal resolutions not attainable with existing optical microscopic techniques. However, currently available GECIs optimized for fluorescence (FL) imaging lack sufficient contrast for OA imaging and respond at wavelengths having limited penetration into the mammalian brain. Here we present an imaging platform capable of rapid assessment and cross-validation between OA and FL responses of sensor proteins expressed in Escherichia coli colonies. The screening system features optimized pulsed light excitation combined with ultrasensitive ultrasound detection to mitigate photobleaching while further allowing the dynamic characterization of calcium ion responses with millisecond precision. Targeted probing of up to six individual colonies per second in both calcium-loaded and calcium-unloaded states was possible with the system. The new platform greatly facilitates optimization of absorption-based labels, thus setting the stage for directed evolution of OA GECIs.

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Short and long-term phototoxicity in cells expressing genetic reporters under nanosecond laser exposure

Cited by 8 publications

References 38 publications

Advanced optoacoustic methods for multiscale imaging of in vivo dynamics

Advanced optoacoustic methods for multiscale imaging of in vivo dynamics

Investigating fibroblast cells under “safe” and “injurious” blue‐light exposure by holographic microscopy

High-Throughput Platform for Optoacoustic Probing of Genetically Encoded Calcium Ion Indicators

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