Three-dimensional optical-transfer-function analysis of fiber-optical two-photon fluorescence microscopy

Gu, Miṅ; Bird, Damian

doi:10.1364/josaa.20.000941

Cited by 10 publications

(6 citation statements)

References 21 publications

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“…Here a and r 0 are the radius of the GRIN rod lens and the fiber mode profile, respectively, is the illumination wavelength, and d is the gap length. As was shown in an earlier study, 14 C 2 ͑l ϭ 0͒ decreases monotonically as A increases. As a result of the balance between A 4 ͓͞1 Ϫ exp͑ϪA͔͒ and C 2 ͑l ϭ 0͒, the signal level given by Eq.…”

Section: Optimization Of a Two-photon Fluorescence Signalsupporting

confidence: 82%

“…The appearance of the maximum signal level when the gap length increases can be understood from the optical transfer function analysis. 14 According to this method, 11,14 the signal level of a thin fluorescence sheet is proportional to the value of the twodimensional in-focus optical transfer function C 2 ͑l͒ at l ϭ 0, where l is the transverse spatial frequency of an object. 11 In the case of the fiber-optic two-photon fluorescence microscope that uses a single-mode fiber coupler and a GRIN rod lens, the signal level can be expressed, if the input power from the Gaussian fiber profile and the power loss by the GRIN rod lens are considered, as…”

Section: Optimization Of a Two-photon Fluorescence Signalmentioning

confidence: 99%

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Characterization of gradient-index lens-fiber spacing toward applications in two-photon fluorescence endoscopy

Gan

2005

Appl. Opt.

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We report on the experimental investigation into the characterization of two-photon fluorescence microscopy based on the separation distance of a single-mode optical fiber coupler and a gradient-index (GRIN) rod lens. The collected two-photon fluorescence signal exhibits a maximum intensity at a defined separation distance (gap length) where the increasing effective excitation numerical aperture is balanced by the decreasing confocal emission collection. A maximum signal is found at gap lengths of approximately 2, 1.25, and 1.75 mm for GRIN lenses with pitches of 0.23, 0.25, and 0.29 wavelength at 830 nm. The maximum two-photon fluorescence signal collected corresponds to a threefold reduction of axial resolution (38.5 microm at 1.25 mm), compared with the maximum resolution (11.6 microm at 5.5 mm), as shown by the three-dimensional imaging of 10 microm beads. These results demonstrate an intrinsic trade-off between signal collection and axial resolution.

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Section: Optimization Of a Two-photon Fluorescence Signalsupporting

confidence: 82%

Section: Optimization Of a Two-photon Fluorescence Signalmentioning

confidence: 99%

Characterization of gradient-index lens-fiber spacing toward applications in two-photon fluorescence endoscopy

Gan

2005

Appl. Opt.

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“…Since the excitation probability for 2P fluorescence varies quadratically with the illuminating intensity, the PSF of the 2P probe also varies as the square of the laser intensity distribution. The latter is determined by a convolution of the fiber output laser intensity distribution and the impulse response of the focusing optics [35]. The present instrument has a non-confocal detection geometry because most of the fluorescence signal is collected by the inner cladding of the fiber.…”

Section: Design Criteria and Performance Of The Endoscopementioning

confidence: 99%

Development of a versatile two-photon endoscope for biological imaging

Zhao

Nakamura

Gordon

2010

Biomed. Opt. Express

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We describe a versatile, catheter-type two-photon probe, designed for in vivo and ex vivo imaging of the aqueous outflow pathway in the eye. The device consists of a silica double cladding fiber used for laser delivery and fluorescence collection, a spiral fiber scanner driven by a miniature piezoelectric tube, and an assembly of three micro-size doublet achromatic lenses used for focusing the laser and collecting the two-photon excitation signal. All the components have a maximum diameter of 2 mm and are enclosed in a length of 12-gauge stainless steel hypodermic tubing having an outer diameter of 2.8 mm. The lateral and axial resolutions of the probe are measured to be 1.5 μm and 9.2 μm, respectively. Different lens configurations and fibers are evaluated by comparing their spatial resolutions and fluorescence signal collection efficiencies. Doublet achromatic lenses and a double cladding fiber with a high inner cladding numerical aperture are found to produce a high signal collection efficiency, which is essential for imaging live tissues. Simple methods for reducing image distortions are demonstrated. Images of human trabecular meshwork tissue are successfully obtained with this miniature two-photon microscope.

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“…The application of this geometry has been extended to SHG imaging to analyse the molecular orientations of structural proteins (Fu et al , 2005c, d). A nonlinear optical microscope based on a fibre coupler exhibits a confocal image nature, improving image resolution (Gu & Bird, 2003; Gu & Fu, 2006). More importantly, an introduction of a double‐clad PCF to TPEF and SHG endoscopy has resulted in significant enhancement of signal level (Fu et al , 2005a).…”

Section: Geometries Of Fibre‐optic Nonlinear Optical Microscopymentioning

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 optical-transfer-function analysis of fiber-optical two-photon fluorescence microscopy

Cited by 10 publications

References 21 publications

Characterization of gradient-index lens-fiber spacing toward applications in two-photon fluorescence endoscopy

Characterization of gradient-index lens-fiber spacing toward applications in two-photon fluorescence endoscopy

Development of a versatile two-photon endoscope for biological imaging

Fibre‐optic nonlinear optical microscopy and endoscopy

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