Wide-field multiphoton imaging of cellular dynamics in thick tissue by temporal focusing and patterned illumination

Therrien, O. D.; Aubé, Benoît; Pagès, Stéphane; Koninck, Paul De; Côté, Daniel

doi:10.1364/boe.2.000696

Cited by 69 publications

(49 citation statements)

References 23 publications

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“…Integrated with patterned illumination, the system could further be applied for functional excitation [17,28,29,31], 3D fabrication [30], and structured illumination purposes [39][40][41][42]. In this paper, we have demonstrated that the AEC would vary with different patterns at the same single spatial frequency and different orientations, and could reach nearly 29 % difference, e. g., from À11 % to + 18 % with 1.09 mm À1 spatial frequency patterns at the 908 and 08 orientations.…”

Section: Discussionmentioning

confidence: 89%

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Chang

Lin

et al. 2017

Journal of Biophotonics

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A developed temporal focusing-based multiphoton excitation microscope (TFMPEM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back-focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPEM can be condensed from 3.0 mm to 1.5 mm for a 50 % improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture: TPEF images of the eosin-stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 mm À1 spatially modulated illumination at 908 (center) and 08 (right) orientations.

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Section: Discussionmentioning

confidence: 89%

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Chang

Lin

et al. 2017

Journal of Biophotonics

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“…Compared to conventional beam scanning multiphoton microscopy, widefield multiphoton microscopy using the temporal focusing technique detects the overall fluorescence and harmonic generation signal of the entire illumination area, which depends on the laser beam spot size and the magnification of the microscope. The advantage of widefield multiphoton microscopy is that less time is required to capture one frame, enabling a fast frame rate for capturing dynamic events [7,8] or fast microfabrication applications [9,10]. With a fast, high-sensitivity camera and an ultrahigh peak power laser, an imaging rate of a few hundred frames per second can be achieved in widefield multiphoton microscopy [11].…”

Section: Introductionmentioning

confidence: 99%

Wavefront sensorless adaptive optics temporal focusing-based multiphoton microscopy

Chang

et al. 2014

Biomed. Opt. Express

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Temporal profile distortions reduce excitation efficiency and image quality in temporal focusing-based multiphoton microscopy. In order to compensate the distortions, a wavefront sensorless adaptive optics system (AOS) was integrated into the microscope. The feedback control signal of the AOS was acquired from local image intensity maximization via a hill-climbing algorithm. The control signal was then utilized to drive a deformable mirror in such a way as to eliminate the distortions. With the AOS correction, not only is the axial excitation symmetrically refocused, but the axial resolution with full two-photon excited fluorescence (TPEF) intensity is also maintained. Hence, the contrast of the TPEF image of a R6G-doped PMMA thin film is enhanced along with a 3.7-fold increase in intensity. Furthermore, the TPEF image quality of 1μm fluorescent beads sealed in agarose gel at different depths is improved.

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“…It has been successfully implemented to image biological samples [3,4], cellular dynamics [5,6], as well as for microfabrication [7][8][9]. TF2p microscopy is, in essence, simply a 4-f pulse shaper proposed by Froehly, Colombeau, and Vampouille [10] and Martinez, Gordon, and Fork [11] with the exception that there is no second grating to recombine the pulse.…”

Section: Introductionmentioning

confidence: 99%

Temporally focused wide-field two-photon microscopy: Paraxial to vectorial

Yew

Sheppard

2013

Opt. Express

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Temporal focusing allows for optically sectioned wide-field microscopy. The optical sectioning arises because this method takes a pulsed input beam, stretches the pulses by diffracting off a grating, and focuses the stretched pulses such that only at the focal plane are the pulses re-compressed. This approach generates nonlinear optical processes at the focal plane and results in depth discrimination. Prior theoretical models of temporal focusing processes approximate the contributions of the different spectral components by their mean. This is valid for longer pulses that have narrower spectral bandwidth but results in a systematic deviation when broad spectrum, femtosecond pulses are used. Further, prior model takes the paraxial approximation but since these pulses are focused with high numerical aperture (NA) objectives, the effects of the vectorial nature of light should be considered. In this paper we present a paraxial and a vector theory of temporal focusing that takes into account the finite spread of the spectrum. Using paraxial theory we arrive at an analytical solution to the electric field at the focus for temporally focused wide-field two-photon (TF2p) microscopy as well as in the case of a spectrally chirped input beam. We find that using paraxial theory while accounting for the broad spectral spread gives results almost twice vector theory. Experiment results agree with predictions of the vector theory giving an axial full-width half maximum (FWHM) of 2.1μm and1.8 μm respectively as long as spectral spread is taken into account. Using our system parameters, the optical sectioning of the TF2p microscope is found to be 8 μm . The optical transfer function (OTF) of a TF2p microscope is also derived and is found to pass a significantly more limited band of axial frequencies than a point scanning two-photon (2p) microscope or a single photon (1p) confocal microscope.

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Wide-field multiphoton imaging of cellular dynamics in thick tissue by temporal focusing and patterned illumination

Cited by 69 publications

References 23 publications

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

Wavefront sensorless adaptive optics temporal focusing-based multiphoton microscopy

Temporally focused wide-field two-photon microscopy: Paraxial to vectorial

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