scholarly journals Fast wavefront shaping for two-photon brain imaging with large field of view correction

2021 ◽  
Author(s):  
Baptiste Blochet ◽  
Walther Akemann ◽  
Sylvain Gigan ◽  
Laurent Bourdieu
Keyword(s):  
Biological Tissues ◽  
Field Of View ◽  
Large Field ◽  
Light Modulation ◽  
Adaptive Optimization ◽  
Wavefront Correction ◽  
Scan Speed ◽  
Acquisition Speed ◽  
Neuron Imaging

In-vivo optical imaging with diffraction-limited resolution deep inside scattering biological tissues is obtained by non-linear fluorescence microscopy. Active compensation of tissue-induced aberrations and light scattering through adaptive wavefront correction further extends depth penetration by restoring high resolution at large depth. However, at large depths those corrections are only valid over a very limited field of view within the angular memory effect. To overcome this limitation, we introduce an acousto-optic light modulation technique for fluorescence imaging with simultaneous wavefront correction at pixel scan speed. Biaxial wavefront corrections are first learned by adaptive optimization at multiple locations in the image field. During image acquisition, the learned corrections are then switched on-the-fly according to the position of the excitation focus during the raster scan. The proposed microscope is applied to in-vivo transcranial neuron imaging and demonstrates correction of skull-induced aberrations and scattering across large fields of view at 40 kHz data acquisition speed.

scholarly journals Large field-of-view incoherent volumetric imaging in living human retina by confocal oblique scanning laser ophthalmoscopy

2021 ◽  
Author(s):  
Wenjun Shao ◽  
Ji Yi
Keyword(s):  
Clinical Care ◽  
Retinal Imaging ◽  
Field Of View ◽  
Scanning Laser ◽  
Large Field ◽  
Scanning Laser Ophthalmoscopy ◽  
Human Retina ◽  
Label Free ◽  
Volumetric Imaging

Three-dimensional (3D) volumetric imaging of the human retina is instrumental to monitor and diagnose blinding conditions. Although coherent retinal imaging is well established by optical coherence tomography, it is still a large void for incoherent volumetric imaging in the human retina. Here, we report confocal oblique scanning laser ophthalmoscopy (CoSLO), to fill that void and harness incoherent optical contrast in 3D. CoSLO uses oblique scanning laser and remote focusing to acquire depth signal in parallel, avoid the lengthy z-stacking, and image a large field of view (FOV). In addition, confocal gating is introduced by a linear sensor array to improve the contrast and resolution. For the first time, we achieved incoherent 3D human retinal imaging with >20° viewing angle within only 5 seconds. The depth resolution is ~45 microns in vivo. We demonstrated label-free incoherent contrast by CoSLO, revealing unique features in the retina. CoSLO will be an important technique for clinical care of retinal conditions and fundamental vision science, by offering unique volumetric incoherent contrasts.

scholarly journals Integrating photoacoustic microscopy, optical coherence tomography, OCT angiography, and fluorescence microscopy for multimodal imaging

2020 ◽  
Vol 245 (4) ◽  
pp. 342-347 ◽  
Author(s):  
Arash Dadkhah ◽  
Shuliang Jiao
Keyword(s):  
Optical Coherence Tomography ◽  
Fluorescence Microscopy ◽  
Multimodal Imaging ◽  
Imaging System ◽  
Optical Resolution ◽  
Biological Tissues ◽  
Large Field ◽  
Optical Coherence ◽  
Photoacoustic Microscopy

We have developed a multimodal imaging system, which integrated optical resolution photoacoustic microscopy, optical coherence tomography, optical coherence tomography angiography, and confocal fluorescence microscopy in one platform. The system is able to image complementary features of a biological sample by combining different contrast mechanisms. We achieved fast imaging and large field of view by combining optical scanning with mechanical scanning, similar to our previous publication. We have demonstrated the capability of the multimodal imaging system by imaging a mouse ear in vivo. Impact statement Photoacoustic microscopy-based multimodal imaging technology can provide high-resolution complementary information for biological tissues in vivo. It will potentially bring significant impact on the research and diagnosis of diseases by providing combined structural and functional information.

Large Field-of-View and Deep Tissue Optical Micro-Imaging Based on Parallel Wavefront Correction Algorithm

2018 ◽  
Vol 45 (12) ◽  
pp. 1207001
Author(s):  
赵琪 Zhao Qi ◽  
石鑫 Shi Xin ◽  
龚薇 Gong Wei ◽  
胡乐佳 Hu Lejia ◽  
郑瑶 Zheng Yao ◽  
...  
Keyword(s):  
Field Of View ◽  
Large Field ◽  
Correction Algorithm ◽  
Deep Tissue ◽  
Wavefront Correction

scholarly journals A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Nicholas James Sofroniew ◽  
Daniel Flickinger ◽  
Jonathan King ◽  
Karel Svoboda
Keyword(s):  
Neural Activity ◽  
Single Cells ◽  
Numerical Aperture ◽  
Random Access ◽  
Field Of View ◽  
Large Field ◽  
Excitation Wavelength ◽  
Scanning System ◽  
Brain Areas

Imaging is used to map activity across populations of neurons. Microscopes with cellular resolution have small (<1 millimeter) fields of view and cannot simultaneously image activity distributed across multiple brain areas. Typical large field of view microscopes do not resolve single cells, especially in the axial dimension. We developed a 2-photon random access mesoscope (2p-RAM) that allows high-resolution imaging anywhere within a volume spanning multiple brain areas (∅ 5 mm x 1 mm cylinder). 2p-RAM resolution is near diffraction limited (lateral, 0.66 μm, axial 4.09 μm at the center; excitation wavelength = 970 nm; numerical aperture = 0.6) over a large range of excitation wavelengths. A fast three-dimensional scanning system allows efficient sampling of neural activity in arbitrary regions of interest across the entire imaging volume. We illustrate the use of the 2p-RAM by imaging neural activity in multiple, non-contiguous brain areas in transgenic mice expressing protein calcium sensors.

Compound Scanning with a Phased Array

1982 ◽  
Vol 4 (2) ◽  
pp. 93-107 ◽  
Author(s):  
David P. Shattuck ◽  
Olaf T. von Ramm
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Phased Array ◽  
Speckle Noise ◽  
Abdominal Imaging ◽  
Field Of View ◽  
Large Field ◽  
Grey Scale ◽  
Computer Controlled

Compound scans made with a dynamically focussed phased array system have been produced in real time. The scanner, intended for abdominal imaging, has a large field of view. The compounding improves the acquisition of echoes from specular targets by changing the orientation of the insonifying beam and also reduces the speckle noise in grey scale images. These gains are achieved while maintaining the high resolution and flexibility of a computer controlled phased array sector scanner. The configuration of the compound scanner is described, and in vivo abdominal scans are presented.

scholarly journals A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging

2016 ◽  
Author(s):  
N. J. Sofroniew ◽  
D. Flickinger ◽  
J. King ◽  
K. Svoboda
Keyword(s):  
Neural Activity ◽  
Single Cells ◽  
Numerical Aperture ◽  
Random Access ◽  
Field Of View ◽  
Large Field ◽  
Excitation Wavelength ◽  
Scanning System ◽  
Brain Areas

AbstractImaging is used to map activity across populations of neurons. Microscopes with cellular resolution have small (< 1 millimeter) fields of view and cannot simultaneously image activity distributed across multiple brain areas. Typical large field of view microscopes do not resolve single cells, especially in the axial dimension. We developed a 2-photon random access mesoscope (2p-RAM) that allows high-resolution imaging anywhere within a volume spanning multiple brain areas (Ø 5 mm × 1 mm cylinder). 2p-RAM resolution is near diffraction limited (lateral, 0.66 μm, axial 4.09 μm at the center; excitation wavelength = 970 nm; numerical aperture = 0.6) over a large range of excitation wavelengths. A fast threedimensional scanning system allows efficient sampling of neural activity in arbitrary regions of interest across the entire imaging volume. We illustrate the use of the 2p-RAM by imaging neural activity in multiple, non-contiguous brain areas in transgenic mice expressing protein calcium sensors.

scholarly journals Detection of cerebral tauopathy in P301L mice using high-resolution large-field multifocal illumination fluorescence microscopy

2020 ◽  
Author(s):  
Ruiqing Ni ◽  
Zhenyue Chen ◽  
Juan A. Gerez ◽  
Gloria Shi ◽  
Quanyu Zhou ◽  
...  
Keyword(s):  
High Resolution ◽  
Fluorescence Microscopy ◽  
Field Of View ◽  
Large Field ◽  
Depth Of Focus ◽  
Microscopy Technique ◽  
Microscopy Techniques ◽  
Small Field Of View

AbstractCurrent intravital microscopy techniques visualize tauopathy with high-resolution, but have a small field-of-view and depth-of-focus. Herein, we report a transcranial detection of tauopathy over the entire cortex of P301L tauopathy mice using large-field multifocal illumination (LMI) fluorescence microscopy technique and luminescent conjugated oligothiophenes. In vitro assays revealed that fluorescent ligand h-FTAA is optimal for in vivo tau imaging, which was confirmed by observing elevated probe retention in the cortex of P301L mice compared to non-transgenic littermates. Immunohistochemical staining further verified the specificity of h-FTAA to detect tauopathy in P301L mice. The new imaging platform can be leveraged in pre-clinical mechanistic studies of tau spreading and clearance as well as longitudinal monitoring of tau targeting therapeutics.

scholarly journals Relationship between simultaneously recorded spiking activity and fluorescence signal in GCaMP6 transgenic mice

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Lawrence Huang ◽  
Peter Ledochowitsch ◽  
Ulf Knoblich ◽  
Jérôme Lecoq ◽  
Gabe J Murphy ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Action Potentials ◽  
Pyramidal Neurons ◽  
Field Of View ◽  
Large Field ◽  
Fluorescence Signal ◽  
Calcium Indicators ◽  
The Relationship ◽  
Imaging Conditions

Fluorescent calcium indicators are often used to investigate neural dynamics, but the relationship between fluorescence and action potentials (APs) remains unclear. Most APs can be detected when the soma almost fills the microscope's field of view, but calcium indicators are often used to image populations of neurons, necessitating a large field of view, generating fewer photons per neuron, and compromising AP detection. Here we characterized the AP-fluorescence transfer function in vivo for 48 layer 2/3 pyramidal neurons in primary visual cortex, with simultaneous calcium imaging and cell-attached recordings from transgenic mice expressing GCaMP6s or GCaMP6f. While most APs were detected under optimal conditions, under conditions typical of population imaging studies only a minority of 1AP and 2AP events were detected (often <10% and ~20-30%, respectively), emphasizing the limits of AP detection under more realistic imaging conditions.

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