Dual-color deep-tissue three-photon microscopy with a multiband infrared laser

Guesmi, Khmaies; Abdeladim, Lamiae; Tozer, Samuel; Mahou, Pierre; Kumamoto, Takuma; Jurkus, Karolis; Rigaud, Philippe; Loulier, Karine; Dray, Nicolas; Georges, Patrick; Hanna, Marc; Livet, Jean; Supatto, Willy; Beaurepaire, Emmanuel; Druon, Frédéric

doi:10.1038/s41377-018-0012-2

Cited by 113 publications

(83 citation statements)

References 33 publications

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“…pulsed lasers typically have pulse duration and repetition rate below 70 fs and 1.25 MHz, [17][18][19][20][21]23 respectively. The Ti:Sapphire laser source in our system is typically used to generate 2P excitation, therefore, it is not optimal for 3P microscopy.…”

Section: Three-photon Trafixmentioning

confidence: 99%

Wide-field multiphoton imaging with TRAFIX

Escobet-Montalbán

Wijesinghe

Chen

et al. 2019

Multiphoton Microscopy in the Biomedical Sciences XIX

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Optical approaches have broadened their impact in recent years with innovations in both wide-field and superresolution imaging, which now underpin biological and medical sciences. Whilst these advances have been remarkable, to date, the ongoing challenge in optical imaging is to penetrate deeper. TRAFIX is an innovative approach that combines temporal focusing illumination with single-pixel detection to obtain wide-field multiphoton images of fluorescent microscopic samples deep through scattering media without correction. It has been shown that it can image through biological samples such as rat brain or human colon tissue up to a depth of seven scattering mean-free-path lengths. Comparisons of TRAFIX with standard point-scanning two-photon microscopy show that the former can yield a five-fold higher signal-to-background ratio while significantly reducing photobleaching of the specimen. Here, we show the first preliminary demonstration of TRAFIX with three-photon excitation imaging dielectric beads. We discuss the advantages of the TRAFIX approach combined with compressive sensing for biomedicine.

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Section: Three-photon Trafixmentioning

confidence: 99%

Wide-field multiphoton imaging with TRAFIX

Escobet-Montalbán

Wijesinghe

Chen

et al. 2019

Multiphoton Microscopy in the Biomedical Sciences XIX

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“…However, tissue scattering (µ s ) is reduced at these wavelengths. Thus, the choice of the optimal fluorophore and excitation wavelength windows will be a trade-off between absorption and tissue scattering (39,40). Finally, thermal damage is a particular concern for deep-tissue imaging because compensating for the exponential excitation losses with increasing depth will expose the tissue surface to very high laser powers that -even unfocused -will be harmful due to surface heat generation.…”

Section: Brain Heating As a Damage Mechanismmentioning

confidence: 99%

Two-photon imaging induces brain heating and calcium microdomain hyperactivity in cortical astrocytes

Schmidt¹,

Oheim²

2018

Preprint

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Unraveling how neural networks process and represent sensory information and how this cellular dynamics instructs behavioral output is a main goal in current neuroscience.Two-photon activation of optogenetic actuators and fluorescence calcium (Ca 2+ ) imaging with genetically encoded Ca 2+ indicators allow, respectively, the all-optical stimulation and readout of activity from genetically identified cell populations. However, these techniques expose the brain to high near-infrared light doses raising the concern of light-induced adverse effects on the biological phenomena being studied. Combing Ca 2+ imaging of GCaMP6f-expressing cortical astrocytes as a sensitive readout for photodamage and an unbiased machine-based event detection, we demonstrate the subtle build-up of aberrant microdomain Ca 2+ signals in fine astroglial processes. Illumination conditions routinely being used in biological twophoton microscopy (920-nm excitation, 100-fs regime, ten mW average power) increased the frequency of microdomain Ca 2+ events, but left their amplitude, area and duration rather unchanged. This increase in local Ca 2+ activity was followed by Ca 2+ transients in the otherwise silent soma. Ca 2+ hyperactivity occurred without overt morphological damage.Surprisingly, at the same average power, continuous-wave 920-nm illumination was as damaging as fs pulses, indicating a linear, heating-mediated (rather than a highly non-linear) damage mechanism. In an astrocyte-specific IP 3 -receptor knock-out mouse (IP 3 R2-KO), Near-infrared light-induced Ca 2+ microdomains signals persisted in the small processes, underpinning their resemblance to physiological IP 3 R2-independent Ca 2+ signals, while somatic activity was abolished. Contrary to what has generally been believed in the field, shorter pulses and lower average power are advantageous to alleviate photodamage and allow for longer useful recording windows. [250/250 words] Schmidt & Oheim (2018) biorXiv Heating, not highly non-linear photodamage is limiting at 920 nm 3 SIGNIFICANCE STATEMENTImaging the fine structure and function of the brain has become possible with two-photon microscopy that uses ultrashort-pulsed infrared laser light for better tissue penetration. The high peak energy of these light pulses has raised concerns about photodamage resulting from multi-photon processes. Here, we show that the time-averaged rather than the peak laser power matters. At wavelengths and with laser powers now commonly used in neuroscience brain damage occurs as a consequence of direct infrared light absorption, i.e., heating. To counteract brain heating we explore a strategy that uses even shorter, more energetic pulses but a lower time-averaged laser power to produce the same image quality while making twophoton microscopy less invasive. [112/120 words] \body

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“…Crystal-based optical parametric oscillators and amplifiers are conventional methods for producing laser pulses near 1700 nm [7,8]. However, the critical alignment of the nonlinear crystal makes the system bulky and sensitive to the environment.…”

Section: Introductionmentioning

confidence: 99%

All-fiber high-power 1700 nm femtosecond laser based on optical parametric chirped-pulse amplification

Qin¹,

Batjargal²,

Cromey³

et al. 2020

Opt. Express

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We present the design and construction of an all-fiber high-power optical parametric chirped-pulse amplifier working at 1700 nm, an important wavelength for bio-photonics and medical treatments. The laser delivers 1.42 W of output average power at 1700 nm, which corresponds to ∼40 nJ pulse energy. The pulse can be de-chirped with a conventional grating pair compressor to ∼450 fs. Furthermore, the laser has a stable performance with relative intensity noise typically below the -130 dBc/Hz level for the idler pulses at 1700 nm from 10kHz to 16.95 MHz, half of the laser repetition rate f /2.

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Dual-color deep-tissue three-photon microscopy with a multiband infrared laser

Cited by 113 publications

References 33 publications

Wide-field multiphoton imaging with TRAFIX

Wide-field multiphoton imaging with TRAFIX

Two-photon imaging induces brain heating and calcium microdomain hyperactivity in cortical astrocytes

All-fiber high-power 1700 nm femtosecond laser based on optical parametric chirped-pulse amplification

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