scholarly journals Simultaneous fluorescence imaging of tilted focal planes at two depths in thick neural tissue: Implementation with remote focus in a widefield electrophysiological microscope

2019 ◽  
Author(s):  
Bruno Lagarde ◽  
Noah Russell ◽  
Elric Esposito ◽  
Laura Desban ◽  
Claire Wyart ◽  
...  
Keyword(s):  
Brain Slice ◽  
Optical Axis ◽  
Image Plane ◽  
Wide Field ◽  
Single Image ◽  
Time Required ◽  
Field Imaging ◽  
Wide Field Imaging ◽  
Neuronal Compartments

AbstractWide-field imaging conventionally results in a single image plane oriented perpendicular to the optical axis. However, in brain slice or in vivo recording, neuronal or circuit morphologies lie in arbitrarily tilted planes. Consequently the spatiotemporal advantages of wide-field non-scanned imaging are lost because of the time required for stepwise focal readjustments to view an entire neuron or network. We describe an application of remote focus that views simultaneously two planes separated by up to 100 µm, each with variable tilt from the conventional image plane. This permits fluorescence detection of ion fluxes or membrane potential across neuronal compartments and their correlation with electrical activity. Further, two fluorophores can be viewed simultaneously in each plane.We show (i) neuronal images tilted to optimise simultaneous aquisition of somatic, dendritic and axonal compartments; (ii) networks viewed simultaneously at 2 depths separated by up to 100 µm, (iii) widefield imaging at 30 Hz of Gcamp5 fluorescence during spontaneous spiking in motoneuron layers of zebrafish spinal cord separated by 30-40 microns.

scholarly journals Protocol for constructing an extensive cranial window utilizing a PEO-CYTOP nanosheet for in vivo wide-field imaging of the mouse brain

2021 ◽  
Vol 2 (2) ◽  
pp. 100542
Author(s):  
Taiga Takahashi ◽  
Hong Zhang ◽  
Kohei Otomo ◽  
Yosuke Okamura ◽  
Tomomi Nemoto
Keyword(s):  
Mouse Brain ◽  
Wide Field ◽  
Cranial Window ◽  
Field Imaging ◽  
Wide Field Imaging

scholarly journals De-scattering with Excitation Patterning enables rapid wide-field imaging through scattering media

2021 ◽  
Vol 7 (28) ◽  
pp. eaay5496
Author(s):  
Cheng Zheng ◽  
Jong Kang Park ◽  
Murat Yildirim ◽  
Josiah R. Boivin ◽  
Yi Xue ◽  
...  
Keyword(s):  
High Resolution ◽  
Structural Features ◽  
Computational Imaging ◽  
Tissue Imaging ◽  
Wide Field ◽  
Scattering Media ◽  
Temporal Focusing ◽  
Field Imaging ◽  
Wide Field Imaging

Nonlinear optical microscopy has enabled in vivo deep tissue imaging on the millimeter scale. A key unmet challenge is its limited throughput especially compared to rapid wide-field modalities that are used ubiquitously in thin specimens. Wide-field imaging methods in tissue specimens have found successes in optically cleared tissues and at shallower depths, but the scattering of emission photons in thick turbid samples severely degrades image quality at the camera. To address this challenge, we introduce a novel technique called De-scattering with Excitation Patterning or “DEEP,” which uses patterned nonlinear excitation followed by computational imaging–assisted wide-field detection. Multiphoton temporal focusing allows high-resolution excitation patterns to be projected deep inside specimen at multiple scattering lengths due to the use of long wavelength light. Computational reconstruction allows high-resolution structural features to be reconstructed from tens to hundreds of DEEP images instead of millions of point-scanning measurements.

Integrated Raman spectroscopy and trimodal wide-field imaging techniques for real-time in vivo tissue Raman measurements at endoscopy

2009 ◽  
Vol 34 (6) ◽  
pp. 758 ◽  
Author(s):  
Zhiwei Huang ◽  
Seng Khoon Teh ◽  
Wei Zheng ◽  
Jianhua Mo ◽  
Kan Lin ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Real Time ◽  
Imaging Techniques ◽  
Wide Field ◽  
Field Imaging ◽  
Wide Field Imaging

scholarly journals Fast algorithms to approximate the position-dependent point spread function responses in radio interferometric wide-field imaging

2020 ◽  
Vol 499 (1) ◽  
pp. 292-303
Author(s):  
M Atemkeng ◽  
O Smirnov ◽  
C Tasse ◽  
G Foster ◽  
S Makhathini
Keyword(s):  
Point Spread Function ◽  
Simulated Data ◽  
High Sensitivity ◽  
Image Plane ◽  
Three Dimensions ◽  
Wide Field ◽  
Point Spread ◽  
Spread Function ◽  
Field Imaging ◽  
Wide Field Imaging

ABSTRACT The desire for wide field of view, large fractional bandwidth, high sensitivity, high spectral and temporal resolution has driven radio interferometry to the point of big data revolution where the data are represented in at least three dimensions with an axis for spectral windows, baselines, sources, etc., where each axis has its own set of subdimensions. The cost associated with storing and handling these data is very large, and therefore several techniques to compress interferometric data and/or speed up processing have been investigated. Unfortunately, averaging-based methods for visibility data compression are detrimental to the data fidelity, since the point spread function (PSF) is position-dependent, that is, distorted and attenuated as a function of distance from the phase centre. The position dependence of the PSF becomes more severe, requiring more PSF computations for wide-field imaging. Deconvolution algorithms must take the distortion into account in the major and minor cycles to properly subtract the PSF and recover the fidelity of the image. This approach is expensive in computation since at each deconvolution iteration a distorted PSF must be computed. We present two algorithms that approximate these position-dependent PSFs with fewer computations. The first algorithm approximates the position-dependent PSFs in the uv-plane and the second algorithm approximates the position-dependent PSFs in the image plane. The proposed algorithms are validated using simulated data from the MeerKAT telescope.

scholarly journals Beam Combination for Wide Field Imaging

1994 ◽  
Vol 158 ◽  
pp. 83-90
Author(s):  
M. Tallon ◽  
I. Tallon-Bosc
Keyword(s):  
Image Plane ◽  
Field Of View ◽  
Combination Method ◽  
Wide Field ◽  
Radio Interferometry ◽  
Beam Combination ◽  
Stellar Interferometer ◽  
Field Imaging ◽  
Wide Field Imaging ◽  
Pupil Plane

Several limitations reduce the field of view in radio-interferometry. With an optical array, two of them can be overcome to some extent according to the beam combination method. A beam combination in the pupil plane can completely overcome one of them. In the image plane, a beam combination obeying the rules of geometrical optics can overcome both limitations in principle, but is difficult to achieve in practice. We discuss particularly the real case of a Michelson Stellar Interferometer where a periscope partially re-introduces these limitations, yielding a trade-off between the extension of the field of view and the use of the periscope.

scholarly journals Camera-based localization microscopy optimized with calibrated structured illumination

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Martin Schmidt ◽  
Adam C. Hundahl ◽  
Henrik Flyvbjerg ◽  
Rodolphe Marie ◽  
Kim I. Mortensen
Keyword(s):  
Data Analysis ◽  
Super Resolution ◽  
Wide Field ◽  
Structured Illumination ◽  
Uniform Illumination ◽  
Localization Microscopy ◽  
Localization And Tracking ◽  
Field Imaging ◽  
Wide Field Imaging ◽  
Localization Information

AbstractUntil very recently, super-resolution localization and tracking of fluorescent particles used camera-based wide-field imaging with uniform illumination. Then it was demonstrated that structured illuminations encode additional localization information in images. The first demonstration of this uses scanning and hence suffers from limited throughput. This limitation was mitigated by fusing camera-based localization with wide-field structured illumination. Current implementations, however, use effectively only half the localization information that they encode in images. Here we demonstrate how all of this information may be exploited by careful calibration of the structured illumination. Our approach achieves maximal resolution for given structured illumination, has a simple data analysis, and applies to any structured illumination in principle. We demonstrate this with an only slightly modified wide-field microscope. Our protocol should boost the emerging field of high-precision localization with structured illumination.

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