Author response: A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging

AbstractImaging the activity of neurons that are widely distributed across brain regions deep in scattering tissue at high speed remains challenging. Here, we introduce an open-source system with Dual Independent Enhanced Scan Engines for Large Field-of-view Two-Photon imaging (Diesel2p). Combining novel optical design, adaptive optics, and temporal multiplexing, the system offers subcellular resolution over a large field-of-view (∼ 25 mm2) with independent scan engines. We demonstrate the flexibility and various use cases of this system for calcium imaging of neurons in the living brain.

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Diesel2p mesoscope with dual independent scan engines for flexible capture of dynamics in distributed neural circuitry

Nature Communications ◽

10.1038/s41467-021-26736-4 ◽

2021 ◽

Vol 12 (1) ◽

Cited By ~ 1

Author(s):

Che-Hang Yu ◽

Jeffrey N. Stirman ◽

Yiyi Yu ◽

Riichiro Hira ◽

Spencer L. Smith

Keyword(s):

Adaptive Optics ◽

Calcium Imaging ◽

High Speed ◽

Optical Design ◽

Neural Circuitry ◽

Brain Regions ◽

Field Of View ◽

Large Field ◽

Two Photon ◽

Subcellular Resolution

AbstractImaging the activity of neurons that are widely distributed across brain regions deep in scattering tissue at high speed remains challenging. Here, we introduce an open-source system with Dual Independent Enhanced Scan Engines for Large field-of-view Two-Photon imaging (Diesel2p). Combining optical design, adaptive optics, and temporal multiplexing, the system offers subcellular resolution over a large field-of-view of ~25 mm2, encompassing distances up to 7 mm, with independent scan engines. We demonstrate the flexibility and various use cases of this system for calcium imaging of neurons in the living brain.

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In vivo imaging of liver injury and regeneration by functional two-photon microscopy

Zeitschrift für Gastroenterologie ◽

10.1055/s-0036-1597342 ◽

2016 ◽

Vol 54 (12) ◽

pp. 1343-1404

Author(s):

A Ghallab ◽

R Reif ◽

R Hassan ◽

AS Seddek ◽

JG Hengstler

Keyword(s):

Liver Injury ◽

In Vivo Imaging ◽

Two Photon Microscopy ◽

Two Photon

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Intravital multiphoton microscopy of dynamic renal processes

AJP Renal Physiology ◽

10.1152/ajprenal.00473.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. F1084-F1089 ◽

Cited By ~ 118

Author(s):

Bruce A. Molitoris ◽

Ruben M. Sandoval

Keyword(s):

Proximal Tubule ◽

Multiphoton Microscopy ◽

Field Of View ◽

Glomerular Permeability ◽

Structural Alterations ◽

Two Photon Microscopy ◽

Two Photon ◽

Dynamic Events ◽

Subcellular Resolution ◽

Functioning Kidney

Recent advances in microscopy and optics, computer sciences, and the available fluorophores used to label molecules of interest have empowered investigators to utilize intravital two-photon microscopy to study the dynamic events within the functioning kidney. This emerging technique enables investigators to follow functional and structural alterations with subcellular resolution within the same field of view over seconds to weeks. This approach invigorates the validity of data and facilitates analysis and interpretation as trends are more readily determined when one is more closely monitoring indicative physiological parameters. Therefore, in this review we emphasize how specific approaches will enable studies into glomerular permeability, proximal tubule endocytosis, and microvascular function within the kidney. We attempt to show how visual data can be quantified, thus allowing enhanced understanding of the process under study. Finally, emphasis is given to the possible future opportunities of this technology and its present limitations.

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Two-Photon Time-Gated In Vivo Imaging of Dihydrolipoic-Acid-Decorated Gold Nanoclusters

Materials ◽

10.3390/ma14247744 ◽

2021 ◽

Vol 14 (24) ◽

pp. 7744

Author(s):

Ye Tian ◽

Ming Wei ◽

Lijun Wang ◽

Yuankai Hong ◽

Dan Luo ◽

...

Keyword(s):

In Vivo Imaging ◽

Gold Nanoclusters ◽

Photon Absorption ◽

Gold Nanocluster ◽

Time Resolved ◽

Dihydrolipoic Acid ◽

Two Photon Absorption ◽

Two Photon ◽

Time Resolved Imaging

Due to the unique advantages of two-photon technology and time-resolved imaging technology in the biomedical field, attention has been paid to them. Gold clusters possess excellent physicochemical properties and low biotoxicity, which make them greatly advantageous in biological imaging, especially for in vivo animal imaging. A gold nanocluster was coupled with dihydrolipoic acid to obtain a functionalized nanoprobe; the material displayed significant features, including a large two-photon absorption cross-section (up to 1.59 × 105 GM) and prolonged fluorescence lifetime (>300 ns). The two-photon and time-resolution techniques were used to perform cell imaging and in vivo imaging.

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In Vivo Imaging of Neurovascular Coupling with Two-Photon Excitation Laser Scanning Microscopy

The Review of Laser Engineering ◽

10.2184/lsj.40.4_230 ◽

2012 ◽

Vol 40 (4) ◽

pp. 230

Author(s):

Kazuto MASAMOTO ◽

Iwao KANNO

Keyword(s):

In Vivo Imaging ◽

Laser Scanning ◽

Neurovascular Coupling ◽

Laser Scanning Microscopy ◽

Scanning Microscopy ◽

Two Photon ◽

Photon Excitation ◽

Excitation Laser ◽

Two Photon Excitation

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

10.1101/2021.08.05.455286 ◽

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.

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