Bessel-droplet foci enable high-resolution and high-contrast volumetric imaging of synapses and circulation in the brain<i>in vivo</i>

Chen, Wei; Zhang, Qinrong; Natan, Ryan G.; Fan, Jianglan; Ji, Na

doi:10.1101/2022.03.05.483143

Cited by 8 publications

(6 citation statements)

References 38 publications

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“…More importantly, owing to the excellent resolution, contrast, and non-diffracting properties of the Bessel droplet, more sample details can be visualized by the Bessel droplet in contrast to the other two modalities. In contrast to the axially complementary Bessel droplets for achieving a continuous DOF, 41 the TEMP technique not only maintains the high-resolution and high-contrast merits but also allows the 3D reconstruction of sample volumes.…”

Section: ■ Discussionmentioning

confidence: 99%

Tomographic-Encoded Multiphoton Microscopy

Dong

Ren

et al. 2022

ACS Photonics

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Axial scanning in multiphoton microscopy (MPM) is typically realized by mechanically shifting either the objective or the sample. However, the scan speed is usually hindered by the mechanical inertia of the bulky mass. Although the extended depth of field provided by the non-diffracting beam allows fast volumetric imaging, it abandons the axial resolution. Here, we demonstrate a novel and powerful tomographic technique using the Bessel droplet in MPM, termed tomographic-encoded multiphoton (TEMP) microscopy, for structural illumination in the axial direction. We show that benefiting from the high-order nonlinear excitation in MPM, the side-lobe cancellation and smaller beam focus of the Bessel droplet realize better image quality. TEMP microscopy allows fast axial scanning, less risks of photodamage and photobleaching, and high-resolution and high-contrast imaging. Furthermore, fewer raw images are required for the 3D image reconstruction. To demonstrate its usability and advantages for scattering tissues and biomedical applications, we showcase TEMP microscopy with highly scattering fluorescence microspheres and mouse brain slice. More details can be visualized by the Bessel droplet compared with the conventional Gaussian and Bessel beam. The depth-resolving performance of the Bessel droplet has an excellent consistency with the Gaussian beam. More importantly, the TEMP technique is an easy-plug-in method for the current microscopy system. TEMP microscopy is promising for fast volumetric multiphoton imaging, especially for highly scattering tissues.

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Section: ■ Discussionmentioning

confidence: 99%

Tomographic-Encoded Multiphoton Microscopy

Dong

Ren

et al. 2022

ACS Photonics

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“…1 , 0. 5 ( ) (1) where z target is the desired extension range and binarizes the normalized 3D 𝑏𝑖𝑛𝑎𝑟𝑖𝑧𝑒 𝐼 𝑥,𝑦,𝑧;𝑝 ( ) 𝑚𝑎𝑥 𝐼 𝑥,𝑦,𝑧;𝑝 ( ) ( ) , 0. 5 ( )…”

Section: Theory and Designmentioning

confidence: 99%

“…Recent work in tabletop multiphoton neural imaging has shown that wavefront engineering strategies using EDoF imaging and adaptive optics are powerful solutions to improve the imaging capability of neural imaging in-vivo [1,2]. To circumvent the need to headfix animals, the open source head-mounted Miniscope platform [3] is widely adopted for enabling high-resolution neural studies in freely behaving animals.…”

Section: Introductionmentioning

confidence: 99%

EDoF-Miniscope: a computational miniscope for extended depth-of-field neural imaging

Greene

Xue²,

Alido

et al. 2023

Adaptive Optics and Wavefront Control for Biological Systems IX

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We demonstrate an extended-depth-of-field miniscope (EDoF-Miniscope) which utilizes an optimized binary diffractive optical element (DOE) to achieve a 2.8x axial elongation in twin foci when integrated on the pupil plane. We optimize our DOE through a genetic algorithm, which utilizes a Fourier optics forward model to consider the native aberrations of the primary gradient refractive index (GRIN) lens, optical property of the submersion media, the geometric effects of the target fluorescent sources and axial intensity loss from tissue scattering to create a robust EDoF. We demonstrate that our platform achieves high contrast signals that can be recovered through a simple filter across 5-μm and 10-μm beads embedded in scattering phantoms, and fixed mouse brain samples.

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“…Previously, several tabletop systems have demonstrated that EDoF imaging techniques can increase the access of neural imaging without the need for costly inverse algorithms 17 – 20 Typically, the EDoF capability is achieved by pupil engineering, which uses a custom component to modify the optical field as it passes through the pupil plane of a microscope. A large family of pupil functions have been shown to achieve an EDoF 19 , 21 …”

Section: Introductionmentioning

confidence: 99%

Pupil engineering for extended depth-of-field imaging in a fluorescence miniscope

et al. 2023

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Significance: Fluorescence head-mounted microscopes, i.e., miniscopes, have emerged as powerful tools to analyze in-vivo neural populations but exhibit a limited depth-of-field (DoF) due to the use of high numerical aperture (NA) gradient refractive index (GRIN) objective lenses.Aim: We present extended depth-of-field (EDoF) miniscope, which integrates an optimized thin and lightweight binary diffractive optical element (DOE) onto the GRIN lens of a miniscope to extend the DoF by 2.8× between twin foci in fixed scattering samples.Approach: We use a genetic algorithm that considers the GRIN lens' aberration and intensity loss from scattering in a Fourier optics-forward model to optimize a DOE and manufacture the DOE through single-step photolithography. We integrate the DOE into EDoF-Miniscope with a lateral accuracy of 70 μm to produce high-contrast signals without compromising the speed, spatial resolution, size, or weight. Results:We characterize the performance of EDoF-Miniscope across 5-and 10-μm fluorescent beads embedded in scattering phantoms and demonstrate that EDoF-Miniscope facilitates deeper interrogations of neuronal populations in a 100-μm-thick mouse brain sample and vessels in a whole mouse brain sample.Conclusions: Built from off-the-shelf components and augmented by a customizable DOE, we expect that this low-cost EDoF-Miniscope may find utility in a wide range of neural recording applications.

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Bessel-droplet foci enable high-resolution and high-contrast volumetric imaging of synapses and circulation in the brainin vivo

Cited by 8 publications

References 38 publications

Tomographic-Encoded Multiphoton Microscopy

Tomographic-Encoded Multiphoton Microscopy

EDoF-Miniscope: a computational miniscope for extended depth-of-field neural imaging

Pupil engineering for extended depth-of-field imaging in a fluorescence miniscope

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