High speed UHR-OCT for in-vivo volumetric imaging of the palisades of Vogt and the cellular structure of the limbal crypts in the healthy and pathological human corneo-scleral limbus (Conference Presentation)

Author(s):  
Kostadinka Bizheva ◽  
Bingyao Tan ◽  
Zohreh Hosseinaee ◽  
Kirsten Carter ◽  
Denise Hileeto ◽  
...  
Author(s):  
Zohreh Hosseinaee ◽  
Bingyao Tan ◽  
Kirsten Carter ◽  
Denise Hileeto ◽  
Luigina Sorbara ◽  
...  

2016 ◽  
Vol 110 (3) ◽  
pp. 165a ◽  
Author(s):  
Simon P. Poland ◽  
James A. Levitt ◽  
Nikola Krstajić ◽  
Ahmet Erdogen ◽  
Richard J. Walker ◽  
...  

2018 ◽  
Author(s):  
Shuting Han ◽  
Weijian Yang ◽  
Rafael Yuste

To capture the emergent properties of neural circuits, high-speed volumetric imaging of neural activity at cellular resolution is desirable. But while conventional two-photon calcium imaging is a powerful tool to study population activity in vivo, it is restrained to two-dimensional planes. Expanding it to 3D while maintaining high spatiotemporal resolution appears necessary. Here, we developed a two-photon microscope with dual-color laser excitation that can image neural activity in a 3D volume. We imaged the neuronal activity of primary visual cortex from awake mice, spanning from L2 to L5 with 10 planes, at a rate of 10 vol/sec, and demonstrated volumetric imaging of L1 long-range PFC projections and L2/3 somatas. Using this method, we map visually-evoked neuronal ensembles in 3D, finding a lack of columnar structure in orientation responses and revealing functional correlations between cortical layers which differ from trial to trial and are missed in sequential imaging. We also reveal functional interactions between presynaptic L1 axons and postsynaptic L2/3 neurons. Volumetric two-photon imaging appears an ideal method for functional connectomics of neural circuits.


2021 ◽  
Author(s):  
Connor James Darling ◽  
Samuel P.X. Davis ◽  
Sunil Kumar ◽  
Paul M.W. French ◽  
James A McGinty

We present a single-shot adaptation of Optical Projection Tomography (OPT) for high-speed volumetric snapshot imaging of dynamic mesoscopic samples. Conventional OPT has been applied to in vivo imaging of animal models such as D. rerio but the sequential acquisition of projection images required for volumetric reconstruction typically requires samples to be immobilised during the acquisition of an OPT data set. We present a proof-of-principle system capable of single-shot imaging of a 1 mm diameter volume, demonstrating camera-limited rates of up to 62.5 volumes/second, which we have applied to 3D imaging of a freely-swimming zebrafish embryo. This is achieved by recording 8 projection views simultaneously on 4 low-cost CMOS cameras. With no stage required to rotate the sample, this single-shot OPT system can be implemented with a component cost of under 5,000GBP. The system design can be adapted to different sized fields of view and may be applied to a broad range of dynamic samples, including fluid dynamics.


Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 634
Author(s):  
Ruheng Shi ◽  
Yuanlong Zhang ◽  
Tiankuang Zhou ◽  
Lingjie Kong

High-speed, optical-sectioning imaging is highly desired in biomedical studies, as most bio-structures and bio-dynamics are in three-dimensions. Compared to point-scanning techniques, line scanning temporal focusing microscopy (LSTFM) is a promising method that can achieve high temporal resolution while maintaining a deep penetration depth. However, the contrast and axial confinement would still be deteriorated in scattering tissue imaging. Here, we propose a HiLo-based LSTFM, utilizing structured illumination to inhibit the fluorescence background and, thus, enhance the image contrast and axial confinement in deep imaging. We demonstrate the superiority of our method by performing volumetric imaging of neurons and dynamical imaging of microglia in mouse brains in vivo.


2019 ◽  
Vol 42 (1) ◽  
pp. 295-313 ◽  
Author(s):  
Elizabeth M.C. Hillman ◽  
Venkatakaushik Voleti ◽  
Wenze Li ◽  
Hang Yu

Light-sheet microscopy is an imaging approach that offers unique advantages for a diverse range of neuroscience applications. Unlike point-scanning techniques such as confocal and two-photon microscopy, light-sheet microscopes illuminate an entire plane of tissue, while imaging this plane onto a camera. Although early implementations of light sheet were optimized for longitudinal imaging of embryonic development in small specimens, emerging implementations are capable of capturing light-sheet images in freely moving, unconstrained specimens and even the intact in vivo mammalian brain. Meanwhile, the unique photobleaching and signal-to-noise benefits afforded by light-sheet microscopy's parallelized detection deliver the ability to perform volumetric imaging at much higher speeds than can be achieved using point scanning. This review describes the basic principles and evolution of light-sheet microscopy, followed by perspectives on emerging applications and opportunities for both imaging large, cleared, and expanded neural tissues and high-speed, functional imaging in vivo.


2018 ◽  
Author(s):  
Mariya Chavarha ◽  
Vincent Villette ◽  
Ivan K. Dimov ◽  
Lagnajeet Pradhan ◽  
Stephen W. Evans ◽  
...  

ABSTRACTImaging of transmembrane voltage deep in brain tissue with cellular resolution has the potential to reveal information processing by neuronal circuits in living animals with minimal perturbation. Multi-photon voltage imaging in vivo, however, is currently limited by speed and sensitivity of both indicators and imaging methods. Here, we report the engineering of an improved genetically encoded voltage indicator, ASAP3, which exhibits up to 51% fluorescence responses in the physiological voltage range, sub-millisecond activation kinetics, and full responsivity under two-photon illumination. We also introduce an ultrafast local volume excitation (ULOVE) two-photon scanning method to sample ASAP3 signals in awake mice at kilohertz rates with increased stability and sensitivity. ASAP3 and ULOVE allowed continuous single-trial tracking of spikes and subthreshold events for minutes in deep locations, with subcellular resolution, and with repeated sampling over multiple days. By imaging voltage in visual cortex neurons, we found evidence for cell type-dependent subthreshold modulation by locomotion. Thus, ASAP3 and ULOVE enable continuous high-speed high-resolution imaging of electrical activity in deeply located genetically defined neurons during awake behavior.


1992 ◽  
Vol 14 (3) ◽  
pp. 213-233 ◽  
Author(s):  
S.W. Smith ◽  
G.E. Trahey ◽  
O.T. von Ramm

The design, fabrication and evaluation of two-dimensional transducer arrays are described for medical ultrasound imaging. A 4 × 32, 2.8 MHz array was developed to use new signal processing techniques for improved B-scan imaging including elevation focusing, phase correction and synthetic aperture imaging. Laboratory measurements from typical array elements showed 50 Ω dB, and − dB pulse-echo angular response of 62°. Simulations of pulse-echo beam plots have shown grating lobes 20 dB below the main lobe at ±7° in the elevation direction. The complete 2-D array has been used for measurements of phase aberrations in breast, and the individual 32 element linear arrays have been used to obtain conventional B-scans. Several 16 × 16 arrays have also been developed for high speed volumetric imaging. These include 96 transmit elements and 32 receive channels. With a λ/4 matching layer, laboratory measurements show 50 Ω insertion loss of −72 dB, − dB fractional bandwidth of 63%, interelement crosstalk of &29 dB and − dB angular response of 25°. Pulse-echo sensitivity was improved by 21 dB through the use of integrated circuit preamplifiers of high impedance mounted in the transducer handle. In vivo cardiac, abdominal, and obstetric B-scans with elevation focusing, as well as high speed C-scans, have been obtained with these 2-D arrays.


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