Three-dimensional autofluorescence spectroscopy of rat skeletal muscle tissue under two-photon excitation

Schilders, S. P.; Gu, Miṅ

doi:10.1364/ao.38.000720

Cited by 32 publications

(25 citation statements)

References 8 publications

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“…2, a and b, left). TPF from the smooth muscle cells can most likely be attributed to NAD(P)H and flavin compounds (Schilders and Gu, 1999). The smooth muscle cells are invested with curved collagen fibers that exhibit SHG signal (Fig.…”

Section: Characterization Of Mpm Signals From Muscular Arteriesmentioning

confidence: 99%

Imaging Coronary Artery Microstructure Using Second-Harmonic and Two-Photon Fluorescence Microscopy

Zoumi

Kassab

et al. 2004

Biophysical Journal

273

246

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The microstructural basis for the mechanical properties of blood vessels has not been directly determined because of the lack of a nondestructive method that yields a three-dimensional view of these vascular wall constituents. Here, we demonstrate that multiphoton microscopy can be used to visualize the microstructural basis of blood vessel mechanical properties, by combining mechanical testing (distension) of excised porcine coronary arteries with simultaneous two-photon excited fluorescence and second-harmonic generation microscopy. Our results show that second-harmonic generation signals derived from collagen can be spectrally isolated from elastin and smooth muscle cell two-photon fluorescence. Two-photon fluorescence signals can be further characterized by emission maxima at 495 nm and 520 nm, corresponding to elastin and cellular contributions, respectively. Two-dimensional reconstructions of spectrally fused images permit high-resolution visualization of collagen and elastin fibrils and smooth muscle cells from intima to adventitia. These structural features are confirmed by coregistration of multiphoton microscopy images with conventional histology. Significant changes in mean fibril thickness and overall wall dimension were observed when comparing no load (zero transmural pressure) and zero-stress conditions to 30 and 180 mmHg distension pressures. Overall, these data suggest that multiphoton microscopy is a highly sensitive and promising technique for studying the morphometric properties of the microstructure of the blood vessel wall.

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Section: Characterization Of Mpm Signals From Muscular Arteriesmentioning

confidence: 99%

Imaging Coronary Artery Microstructure Using Second-Harmonic and Two-Photon Fluorescence Microscopy

Zoumi

Kassab

et al. 2004

Biophysical Journal

273

246

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“…5 For 1p excitation, an Ar ion laser at a wavelength of 488 nm was used. A Spectra-Physics ultrashort pulsed laser ͑Tsunami͒ with a pulse width of 80 fs was employed for 2p excitation at 800 nm wavelength.…”

Section: ͓S0003-6951͑00͒04536-8͔mentioning

confidence: 99%

“…A Spectra-Physics ultrashort pulsed laser ͑Tsunami͒ with a pulse width of 80 fs was employed for 2p excitation at 800 nm wavelength. 5 To avoid the effect of refractive-index mismatching between the turbid medium and the cover glass of the cell, a water-immersion objective ͑Olympus UplanApo 60ϫ, ϱ/1.13-0.21, numerical apertureϭ1.2, working distanceϭ250 m) was used. In the case of 1p excitation, a pinhole 300 m ͑optical unit ϳ3) in diameter was placed in front of the detector to produce an optical sectioning effect with strength similar to that under 2p excitation without using a pinhole.…”

Section: ͓S0003-6951͑00͒04536-8͔mentioning

confidence: 99%

Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Gan

Kisteman

et al. 2000

Applied Physics Letters

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We show, both theoretically and experimentally, that for a turbid tissue medium where Mie scattering is dominant, multiple scattering not only reduces the illumination power in the forward direction but also exhibits an anisotropic distribution of scattered photons. Thus, a signal level under two-photon excitation drops much faster than that under single-photon excitation although image resolution is much higher in the former case. As a result, the penetration depth under two-photon excitation is limited by the strength of two-photon fluorescence and is not necessarily larger than that under single-photon excitation.

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“…All these nonlinear optical imaging techniques offer unique approaches to visualize 3D tissue structures with high‐resolution and noninvasive means. In particular, the combination of TPEF and SHG has stimulated new insights and information about the cancer research (Schilders & Gu, 1999; Xu et al ., 2000; Deng et al , 2002a; Yeh et al , 2002; Zoumi et al , 2002, 2004; Condeelis & Segall, 2003; Zipfel et al , 2003b). For example, TPEF from intrinsic fluorophores (such as NADH and Flavins) and SHG from extracellular matrix (such as collagen and microtubules) provide the complementary information on general tissue morphology, redox state and fibril orientation (Zipfel et al , 2003b).…”

Section: Nonlinear Optical Imaging Modalitiesmentioning

confidence: 99%

Fibre‐optic nonlinear optical microscopy and endoscopy

2007

Journal of Microscopy

103

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Summary Nonlinear optical microscopy has been an indispensable laboratory tool of high‐resolution imaging in thick tissue and live animals. Rapid developments of fibre‐optic components in terms of growing functionality and decreasing size provide enormous opportunities for innovations in nonlinear optical microscopy. Fibre‐based nonlinear optical endoscopy is the sole instrumentation to permit the cellular imaging within hollow tissue tracts or solid organs that are inaccessible to a conventional optical microscope. This article reviews the current development of fibre‐optic nonlinear optical microscopy and endoscopy, which includes crucial technologies for miniaturized nonlinear optical microscopy and their embodiments of endoscopic systems. A particular attention is given to several classes of photonic crystal fibres that have been applied to nonlinear optical microscopy due to their unique properties for ultrashort pulse delivery and signal collection. Furthermore, fibre‐optic nonlinear optical imaging systems can be classified into portable microscopes suitable for imaging behaving animals, rigid endoscopes that allow for deep tissue imaging with minimally invasive manners, and flexible endoscopes enabling imaging of internal organs. Fibre‐optic nonlinear optical endoscopy is coming of age and a paradigm shift leading to optical microscope tools for early cancer detection and minimally invasive surgery.

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Three-dimensional autofluorescence spectroscopy of rat skeletal muscle tissue under two-photon excitation

Cited by 32 publications

References 8 publications

Imaging Coronary Artery Microstructure Using Second-Harmonic and Two-Photon Fluorescence Microscopy

Imaging Coronary Artery Microstructure Using Second-Harmonic and Two-Photon Fluorescence Microscopy

Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Fibre‐optic nonlinear optical microscopy and endoscopy

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