Gaussian beam in two-photon fluorescence imaging of rat brain microvessel

Shi, Lingyan; Rodríguez-Contreras, Adrián; Alfano, R. R.

doi:10.1117/1.jbo.19.12.126006

Cited by 11 publications

(7 citation statements)

References 13 publications

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“…In the absence of SWIR excitation the signal scaled quadratically with the NIR power (green curve in Fig. 3(b)), which confirms D-2PE of the sample with 825nm excitation [41]. With both lasers illuminating the sample, the fluorescence signal increased with an increase in power of each beam.…”

Section: Power Dependence Of Nd-2pe and D-2pe On Nir And Swir Beamsupporting

confidence: 52%

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy

Yang¹,

Abashin²,

Saisan³

et al. 2016

Opt. Express

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Non-degenerate 2-photon excitation (ND-2PE) of a fluorophore with two laser beams of different photon energies offers an independent degree of freedom in tuning of the photon flux for each beam. This feature takes advantage of the infrared wavelengths used in degenerate 3-photon excitation (D-3PE) microscopy to achieve increased penetration depths, while preserving a relatively high 2-photon excitation cross section in comparison to that of D-3PE. Here, using spatially and temporally aligned Ti:Sapphire laser and optical parametric oscillator beams operating at near infrared (NIR) and short-wavelength infrared (SWIR) optical frequencies, we employ ND-2PE and provide a practical demonstration that a constant fluorophore emission intensity is achievable deeper into a scattering medium using ND-2PE as compared to the commonly used degenerate 2-photon excitation (D-2PE). References and links1. K. Svoboda and R. Yasuda, "Principles of two-photon excitation microscopy and its applications to neuroscience," Neuron 50(6), 823-839 (2006). 2. J. N. Kerr and W. Denk, "Imaging in vivo: watching the brain in action," Nat. Rev. Neurosci. 9(3), 195-205 (2008). 3. W. Denk and K. Svoboda, "Photon upmanship: why multiphoton imaging is more than a gimmick," Neuron 18(3), 351-357 (1997). 4. F. Helmchen and W. Denk, "Deep tissue two-photon microscopy," Nat. Methods 2(12), 932-940 (2005 11. D. Kobat, N. G. Horton, and C. Xu, "In vivo two-photon microscopy to 1.6-mm depth in mouse cortex," J. Biomed. Opt. 16(10), 106014 (2011). 12. R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, "Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser," Sci. Rep. 3, 1014 (2013). 13. P. Theer and W. Denk, "On the fundamental imaging-depth limit in two-photon microscopy," J. Opt. Soc. Am.A adaptive optical recovery of optimal resolution over large volumes," Nat. Methods 11(6), 625-628 (2014).

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Section: Power Dependence Of Nd-2pe and D-2pe On Nir And Swir Beamsupporting

confidence: 52%

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy

Yang¹,

Abashin²,

Saisan³

et al. 2016

Opt. Express

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“…The imaging resolutions of moxifloxacin-based 3P imaging using 1030 nm excitation wavelength were similar to those of moxifloxacin-based 2P imaging using 780 nm excitation wavelength. The Rayleigh length of 3P excitation is smaller than that of 2P excitation, if the same excitation wavelength is used 20 . However, 3P excitation usually requires excitation wavelengths 1.5 times longer than those for 2P excitation for the same fluorophores.…”

Section: Discussionmentioning

confidence: 99%

Three-photon tissue imaging using moxifloxacin

Lee

Wang

et al. 2018

Sci Rep

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Moxifloxacin is an antibiotic used in clinics and has recently been used as a clinically compatible cell-labeling agent for two-photon (2P) imaging. Although 2P imaging with moxifloxacin labeling visualized cells inside tissues using enhanced fluorescence, the imaging depth was quite limited because of the relatively short excitation wavelength (<800 nm) used. In this study, the feasibility of three-photon (3P) excitation of moxifloxacin using a longer excitation wavelength and moxifloxacin-based 3P imaging were tested to increase the imaging depth. Moxifloxacin fluorescence via 3P excitation was detected at a >1000 nm excitation wavelength. After obtaining the excitation and emission spectra of moxifloxacin, moxifloxacin-based 3P imaging was applied to ex vivo mouse bladder and ex vivo mouse small intestine tissues and compared with moxifloxacin-based 2P imaging by switching the excitation wavelength of a Ti:sapphire oscillator between near 1030 and 780 nm. Both moxifloxacin-based 2P and 3P imaging visualized cellular structures in the tissues via moxifloxacin labeling, but the image contrast was better with 3P imaging than with 2P imaging at the same imaging depths. The imaging speed and imaging depth of moxifloxacin-based 3P imaging using a Ti:sapphire oscillator were limited by insufficient excitation power. Therefore, we constructed a new system for moxifloxacin-based 3P imaging using a high-energy Yb fiber laser at 1030 nm and used it for in vivo deep tissue imaging of a mouse small intestine. Moxifloxacin-based 3P imaging could be useful for clinical applications with enhanced imaging depth.

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“…Restrictive motion causes less non radiative pathways and longer lifetime. Critical optical properties of a Gaussian laser beam in two‐photon or multiphoton fluorescence imaging have been theoretically analyzed in ref .…”

Section: Methodsmentioning

confidence: 99%

Alzheimer mouse brain tissue measured by time resolved fluorescence spectroscopy using single‐ and multi‐photon excitation of label free native molecules

Das

Shi

Budansky

et al. 2017

Journal of Biophotonics

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Time resolved spectroscopic measurements with singlephoton and multi-photon excitation of native molecules were performed ex vivo on brain tissues from an Alzheimer's disease (AD) and a wild type (WT) mouse model using a streak camera. The fluorescence decay times of native NADH and FAD show a longer relaxation time in AD than in WT tissue, suggesting less nonradiative processes in AD. The longer emission time of AD may be attributed to the coupling of the key native building block molecules to the amyloid-tau and/or to the caging of the native fluorophores by the deposition of amyloid-beta or tau plaques and neurofibrillary tangles that affect the local non-radiative interactions.

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Gaussian beam in two-photon fluorescence imaging of rat brain microvessel

Cited by 11 publications

References 13 publications

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy

Three-photon tissue imaging using moxifloxacin

Alzheimer mouse brain tissue measured by time resolved fluorescence spectroscopy using single‐ and multi‐photon excitation of label free native molecules

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