Rapid high resolution 3D imaging of expanded biological specimens with lattice light sheet microscopy

Tsai, Yun-Chi; Tang, Wei-Chun; Low, Christine Siok Lan; Liu, Yen-Ting; Wu, Jyun-Sian; Lee, Po-Yi; Chen, Lindsay Quinn; Lin, Yi-Ling; Kanchanawong, Pakorn; Gao, Liang; Chen, Bi-Chang

doi:10.1016/j.ymeth.2019.04.006

Cited by 23 publications

(19 citation statements)

References 26 publications

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“…LSFM offers optical sectioning capability and allows for imaging of biological samples with reduced background and photobleaching [ 75 ]. An improvement in image contrast and resolution was realized with Lattice light sheet microscopy (LLSM), which appears to be an excellent tool in high resolution 3D imaging of fixed tissue up to 20 µm thick [ 76 ] or in expanded biological samples [ 77 ]. A combination of single-molecule localization microscopy with light sheet illumination may allow extremely rapid 3D super-resolution microscopy [ 78 ].…”

Section: Discussionmentioning

confidence: 99%

Comparison of Multiscale Imaging Methods for Brain Research

Tröger

Hoischen

Perner

et al. 2020

Cells

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A major challenge in neuroscience is how to study structural alterations in the brain. Even small changes in synaptic composition could have severe outcomes for body functions. Many neuropathological diseases are attributable to disorganization of particular synaptic proteins. Yet, to detect and comprehensively describe and evaluate such often rather subtle deviations from the normal physiological status in a detailed and quantitative manner is very challenging. Here, we have compared side-by-side several commercially available light microscopes for their suitability in visualizing synaptic components in larger parts of the brain at low resolution, at extended resolution as well as at super-resolution. Microscopic technologies included stereo, widefield, deconvolution, confocal, and super-resolution set-ups. We also analyzed the impact of adaptive optics, a motorized objective correction collar and CUDA graphics card technology on imaging quality and acquisition speed. Our observations evaluate a basic set of techniques, which allow for multi-color brain imaging from centimeter to nanometer scales. The comparative multi-modal strategy we established can be used as a guide for researchers to select the most appropriate light microscopy method in addressing specific questions in brain research, and we also give insights into recent developments such as optical aberration corrections.

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Section: Discussionmentioning

confidence: 99%

Comparison of Multiscale Imaging Methods for Brain Research

Tröger

Hoischen

Perner

et al. 2020

Cells

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“…Seven-day-old WT, Clc-k2–null, and DKO kidneys were optically cleared and expanded before incubation in anti-Nkcc2 antibody (AB2281). The harvested and perfused 7-dayold kidneys were preincubated in gelling solution overnight at 4°C ( 34 ) and the solution was replaced with fresh gelling solution on the second day, followed by incubation at 37°C for 2 hours in a humidified gelation chamber. After the gel polymerization, excess gel around the samples was removed, and the samples were denatured with SDS at 70°C with gentle shaking for 2 days ( 35 ).…”

Section: Methodsmentioning

confidence: 99%

Impairment in renal medulla development underlies salt wasting in Clc-k2 channel deficiency

Lin¹,

Chen²,

Tian³

et al. 2021

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The prevailing view is that ClC-Ka chloride channel (mouse Clc-k1) functions in thin ascending limb for urine concentration, whereas ClC-Kb (mouse Clc-k2) in thick ascending limb (TAL) for salt reabsorption, respectively. Mutations of ClC-Kb cause classic Bartter syndrome with renal salt wasting with onset from perinatal to adolescent.We study the roles of Clc-k channels in perinatal mouse kidneys using constitutive or inducible kidney-specific gene ablation and 2-D and advanced 3-D imaging of optically cleared kidneys. We show that Clc-k1 and -k2 are broadly expressed and colocalized in perinatal kidneys. Deletion of Clc-k1 and -k2 reveals that both participate in NKCC2-and NCC-mediated NaCl reabsorption in neonatal kidneys. Embryonic deletion of Clc-k2 causes tubular injury and impairs renal medulla and TAL development. Inducible deletion of Clc-k2 begins after medulla maturation produces mild salt wasting resulting from reduced NCC activity. Thus, both Clc-k1 and -k2 contribute to salt reabsorption in TAL and DCT in neonates, potentially explaining less severe phenotypes in classic Bartter. As opposed to the current understanding that salt wasting in adult Bartter patients is due to Clc-k2 deficiency in adult TAL, our results suggest that it is mainly originated from medulla and TAL defects during development. Table 2. Plasma and urine biochemistries of 8-week-old Clc-k1-null and Clc-k2-null mice WT Clc-k1 -/-WT Clc-k2 -/-Plasma BUN (mg/dL) 29.7±1.6 26.6±2.0 30.4±1.7 63.6±3.0** Creatinine (mg/dL) 0.25±0.03 0.29±0.03 0.26±0.01 0.39±0.03** Table 3 Plasma and urine biochemistries in 10-week-old inducible Clc-k2 deficient mice.

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“…2 ) [ 45 ]. Optical imaging has witnessed multiple technical innovations (see Table 1 ) [ 45 – 47 ] and is continuously empowered by structured light illumination [ 48 ], digital scanned LSFM [ 49 ], Lattice LSFM [ 50 , 51 ], 2P-LSFM [ 52 ], etc. Many other techniques are reported as well, for instance, to speed up via the extended depth of field microscopy (EDOF) [ 53 – 55 ] or the customized chips [ 56 – 58 ], to improve the spatial resolution via the acousto-optic modulators or spatial light modulators [ 59 , 60 ], to improve the axial accuracy via the depth sensors [ 61 ], to compensate the brain movements with the acousto-optic lens (AOL) 3D random-access pointing and scanning [ 62 ].…”

Section: Optical Microscopymentioning

confidence: 99%

Smart imaging to empower brain-wide neuroscience at single-cell levels

et al. 2022

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A deep understanding of the neuronal connectivity and networks with detailed cell typing across brain regions is necessary to unravel the mechanisms behind the emotional and memorial functions as well as to find the treatment of brain impairment. Brain-wide imaging with single-cell resolution provides unique advantages to access morphological features of a neuron and to investigate the connectivity of neuron networks, which has led to exciting discoveries over the past years based on animal models, such as rodents. Nonetheless, high-throughput systems are in urgent demand to support studies of neural morphologies at larger scale and more detailed level, as well as to enable research on non-human primates (NHP) and human brains. The advances in artificial intelligence (AI) and computational resources bring great opportunity to ‘smart’ imaging systems, i.e., to automate, speed up, optimize and upgrade the imaging systems with AI and computational strategies. In this light, we review the important computational techniques that can support smart systems in brain-wide imaging at single-cell resolution.

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Rapid high resolution 3D imaging of expanded biological specimens with lattice light sheet microscopy

Cited by 23 publications

References 26 publications

Comparison of Multiscale Imaging Methods for Brain Research

Comparison of Multiscale Imaging Methods for Brain Research

Impairment in renal medulla development underlies salt wasting in Clc-k2 channel deficiency

Smart imaging to empower brain-wide neuroscience at single-cell levels

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