Two-photon bioimaging with picosecond optical pulses from a semiconductor laser

Yokoyama, Hiroyuki; Guo, Haotian; Yoda, Takuya; Takashima, Keijiro; Sato, Kimitoshi; Taniguchi, Hiroki; Ito, Hiromasa

doi:10.1364/oe.14.003467

Cited by 65 publications

(38 citation statements)

References 9 publications

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“…Here, the peak-power of the laser is represented as average peak pulse repi P P f τ = , then Na is rewritten as peak average Na P P ∝ This relationship, in which Na is proportional to the product of peak P and average P , can be expected to hold true for both picosecond and femtosecond pulse light sources, as long as the peak power and average power are held constant. Our results demonstrate the utility of a picosecond pulse light source with high peak-power and high average power with low repetition rate for two-photon microscopy [11,13,18]. Actually, Konig's group previously reported that two-photon microscopy using a picosecond laser light source is much less harmful to living cells than a femtosecond laser [19].…”

Section: Resultssupporting

confidence: 58%

“…A picosecond pulse laser with a high repetition rate has been used to successfully excite fluorescent materials and achieve biological imaging [17]. In addition, a highly amplified picosecond pulse enabled us to visualize biological specimens and hippocampal neurons in the mouse brain in vivo [11,13,18]. Here, the number of photons absorbed per fluorophore per second Na [16] increases with the pulse width τ, the square of the peak power, and the repetition frequency as follows: 2 peak pulse repi Na P f τ ∝ Therefore, even with long pulse widths (up to a few picoseconds), we can improve the twophoton fluorescence intensity by decreasing the pulse repetition rate and amplifying the picosecond pulse by using a PCF to 6.7 kW under the objective lens.…”

Section: Resultsmentioning

confidence: 99%

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In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gain-switched laser diode

Kawakami¹,

Sawada²,

Kusama³

et al. 2015

Biomed. Opt. Express

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Abstract:In vivo two-photon microscopy is an advantageous technique for observing the mouse brain at high resolution. In this study, we developed a two-photon microscopy method that uses a 1064-nm gain-switched laser diode-based light source with average power above 4 W, pulse width of 7.5-picosecond, repetition rate of 10-MHz, and a high-sensitivity photomultiplier tube. Using this newly developed two-photon microscope for in vivo imaging, we were able to successfully image hippocampal neurons in the dentate gyrus and obtain panoramic views of CA1 pyramidal neurons and cerebral cortex, regardless of age of the mouse. Fine dendrites in hippocampal CA1 could be imaged with a high peak-signal-tobackground ratio that could not be achieved by titanium sapphire laser excitation. Finally, our system achieved multicolor imaging with neurons and blood vessels in the hippocampal region in vivo. These results indicate that our two-photon microscopy system is suitable for investigations of various neural functions, including the morphological changes undergone by neurons during physiological phenomena.

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Section: Resultssupporting

confidence: 58%

Section: Resultsmentioning

confidence: 99%

In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gain-switched laser diode

Kawakami¹,

Sawada²,

Kusama³

et al. 2015

Biomed. Opt. Express

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“…In order to quantify the suitability of the developed system, the two-photon induced time-averaged fluorescent intensity represents a suitable figure-of-merit (FOM) and can be estimated from P peak × P avg . The calculation arises from nonlinear excitation experiments as the detected signal level from a two-photon process is proportional to this FOM [73]. Semiconductor laser diode systems with amplification schemes have been already successfully demonstrated as light sources for nonlinear microscopy applications [67,74,75].…”

Section: Ec-mlqdl With Postamplification By Tapered Qd-soamentioning

confidence: 99%

Ultra‐Short‐Pulse QD Edge‐Emitting Lasers

Breuer

Syvridis

Рафаилов

2014

The Physics and Engineering of Compact Quantum Dot‐based Lasers for Biophotonics

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IntroductionHigh-power, electrically pumped edge-emitting monolithic semiconductor lasers based on InAs/GaAs quantum dots (QDs) offer significant advantages including ultra-short pulse generation due to ultrafast gain and absorption recovery [1, 2] and emission energies in the near-infrared spectral range [3]. Owing to their inherent inhomogeneous QD size distribution, a broadening of gain and absorption profile is realized, leading to broadband wavelengths emission. This inhomogeneous broadening together with ultrafast carrier dynamics is particularly advantageous for ultrafast applications including the generation, propagation, and amplification of subpicosecond optical pulses with high pulse peak power [4] and the generation of spectrally tunable emission [5,6]. The reduced density of states allows for the exploitation of QD materials in harnessing spectral versatility in novel dual-state emission. The spectral coverage of these QD lasers perfectly matches a wide range of relevant biophotonic applications including nonlinear excitation imaging techniques. The unique combination of efficient light generation, ultrafast carrier dynamics, and broadband gain bandwidth can deliver exclusive advantages in ultrafast microelectronic scale components, thereby stimulating major advances in ultrafast science and technology.In order to enable initial device design and optimization of novel pulse generating QD lasers and amplifiers, a new and efficient theoretical framework is presented as briefly introduced in Section 2.2. This framework takes into account specifically fundamental aspects associated with QD gain materials and allows for modeling of both continuous-wave (CW) emission and ultra-short pulse generation via passive mode-locking (PML) and subsequent amplification. The obtained understanding is applied to technologically realize and optimize QD laser and amplifier structures with particularly broad gain bandwidth enabling spectral versatility through broadband tunability of CW-emission generated in the visible wavelength range by second-harmonic-generation (SHG). This broad wavelength range is accessible only by the QD features as presented in Section 2.3. Novel functionalities based on the broad gain bandwidth and sophisticated QD laser design are explored inThe Physics and Engineering of Compact Quantum Dot-based Lasers for Biophotonics, First Edition. Edited by Edik U. Rafailov.

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“…Another trend towards practical TPM system is the development of compact and turn-key light sources. The emerging compact laser systems are based on semiconductor laser-diode (LD) systems [15,27,[51][52][53]59,60] , optical fiber laser systems [57,58,61] , and even chip-size devices [62] . LD has been used as a reliable, low-cost, highperformance light source in scientific research, optical communication, andinformation technologies.…”

Section: Laser Systemmentioning

confidence: 99%

Recent advances in two-photon imaging: technology developments and biomedical applications

Chen¹,

Guo²,

Gong³

et al. 2013

中国光学快报

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In the past two decades, two-photon microscopy (TPM) transforms biomedicalresearch, allowing nondestructive high-resolution fluorescent molecular imaging and label-free imaging in vivo and in real time.Here we review the recent advances of TPM technologyincluding novel laser sources, new image acquisition paradiams, and microendoscopicimaging systems. Then, we survey the capabilities of TPM imagingof biological relevant molecules such as nicotinamideadenine dinucleotide (NADH), flavin adenine dinucleotide (FAD),and reactive oxygen species (ROS). Biomedical applications of TPM in neuroscience and cancer detection are demonstrated.

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Two-photon bioimaging with picosecond optical pulses from a semiconductor laser

Cited by 65 publications

References 9 publications

In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gain-switched laser diode

In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gain-switched laser diode

Ultra‐Short‐Pulse QD Edge‐Emitting Lasers

Recent advances in two-photon imaging: technology developments and biomedical applications

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