High axial resolution imaging system for large volume tissues using combination of inclined selective plane illumination and mechanical sectioning

Zhang, Qi; Yang, Xiong; Hu, Qinglei; Bai, Ke; Yin, Fang‐Fang; Li, Ning; Gang, Yadong; Wang, Xiaojun; Zeng, Shaoqun

doi:10.1364/boe.8.005767

Cited by 6 publications

(6 citation statements)

References 30 publications

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“…2). On the other hand, the measured axial resolution [ 31 ] of the system is 5 µm, which is limited by the beam width of the illumination. This resolution should be improved if morphological analysis of fine structures, such as spines and single axonal fibers, is desired.…”

Section: Discussionmentioning

confidence: 99%

Scalable volumetric imaging for ultrahigh-speed brain mapping at synaptic resolution

Wang

Zhu

Ding

et al. 2019

National Science Review

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The speed of high-resolution optical imaging has been a rate-limiting factor for meso-scale mapping of brain structures and functional circuits, which is of fundamental importance for neuroscience research. Here, we describe a new microscopy method of Volumetric Imaging with Synchronized on-the-fly-scan and Readout (VISoR) for high-throughput, high-quality brain mapping. Combining synchronized scanning beam illumination and oblique imaging over cleared tissue sections in smooth motion, the VISoR system effectively eliminates motion blur to obtain undistorted images. By continuously imaging moving samples without stopping, the system achieves high-speed 3D image acquisition of an entire mouse brain within 1.5 hours, at a resolution capable of visualizing synaptic spines. A pipeline is developed for sample preparation, imaging, 3D image reconstruction and quantification. Our approach is compatible with immunofluorescence methods, enabling flexible cell-type specific brain mapping and is readily scalable for large biological samples such as primate brains. Using this system, we examined behaviorally relevant whole-brain neuronal activation in 16 c-Fos-shEGFP mice under resting or forced swimming conditions. Our results indicate the involvement of multiple subcortical areas in stress response. Intriguingly, neuronal activation in these areas exhibits striking individual variability among different animals, suggesting the necessity of sufficient cohort size for such studies.

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Section: Discussionmentioning

confidence: 99%

Scalable volumetric imaging for ultrahigh-speed brain mapping at synaptic resolution

Wang

Zhu

Ding

et al. 2019

National Science Review

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“…The collagen layer in the conventional scaffold was not visible using a 30x objective (N.A. = 1.05) which has a Z-resolution of 900 nm [71].…”

Section: Methodsmentioning

confidence: 99%

Molecular transport through primary human small intestinal monolayers by culture on a collagen scaffold with a gradient of chemical cross-linking

et al. 2019

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Background: The luminal surface of the small intestine is composed of a monolayer of cells overlying a lamina propria comprised of extracellular matrix (ECM) proteins. The ECM provides a porous substrate critical for nutrient exchange and cellular adhesion. The enterocytes within the epithelial monolayer possess proteins such as transporters, carriers, pumps and channels that participate in the movement of drugs, metabolites, ions and amino acids and whose function can be regulated or altered by the properties of the ECM. Here, we characterized expression and function of proteins involved in transport across the human small intestinal epithelium grown on two different culture platforms. One strategy employs a conventional scaffolding method comprised of a thin ECM film overlaying a porous membrane while the other utilizes a thick ECM hydrogel placed on a porous membrane. The thick hydrogel possesses a gradient of chemical cross-linking along its length to provide a softer substrate than that of the ECM film-coated membrane while maintaining mechanical stability. Results: The monolayers on both platforms possessed goblet cells and abundant enterocytes and were impermeable to Lucifer yellow and fluorescein-dextran (70 kD) indicating high barrier integrity. Multiple transporter proteins were present in both primary-cell culture formats at levels similar to those present in freshly isolated crypts/villi; however, expression of breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the monolayers on the conventional scaffold was substantially less than that on the gradient cross-linked scaffold and freshly isolated crypts/villi. Monolayers on the conventional scaffold failed to transport the BCRP substrate prazosin while cells on the gradient cross-linked scaffold successfully transported this drug to better mimic the properties of in vivo small intestine. Conclusions:The results of this comparison highlight the need to create in vitro intestinal transport platforms whose characteristics mimic the in vivo lamina propria in order to accurately recapitulate epithelial function.

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“…It is also difficult to diagnose diseases with ground-glass shadows or without a bronchial pathway through imaging. The resolution of confocal microscopy ( 26 , 30 ) can reach 1 µm, and the nucleus can be clearly resolved; however, the field of vision is very small, so it may take a long time to find the lesion site.…”

Section: Discussionmentioning

confidence: 99%

Comparison of the Detection Rates of Different Diagnostic Methods for Primary Peripheral Lung Cancer

Sun

Qin

et al. 2022

Front. Oncol.

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ObjectiveThe present study aims to compare the detection rates of different diagnostic methods for primary peripheral lung cancer (PPLC).MethodsThe detection rate and patient information were collected from a total of 359 cases of PPLC or a suspected diagnosis of lung cancer; among these, 186 cases were simultaneously treated with fibreoptic bronchoscopy, brush inspection and flush inspection, and 173 cases underwent a computed tomography (CT)-guided percutaneous lung biopsy (PNB). The positive detection rates of the different methods were compared.ResultsIn the detection of peripheral lesions (diameter of <5 cm), the CT-PNB had the significantly highest detection rate, followed by the combined basic method (fibreoptic bronchoscopy + brushing + flushing). The independent use of the three basic sampling methods showed a significantly lower detection rate compared with the combined use.ConclusionIn the diagnosis of peripheral lung cancer, the CT-PNB had the best detection rate; hence, it could be used in clinical practice for the diagnosis of such lesions.

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High axial resolution imaging system for large volume tissues using combination of inclined selective plane illumination and mechanical sectioning

Cited by 6 publications

References 30 publications

Scalable volumetric imaging for ultrahigh-speed brain mapping at synaptic resolution

Scalable volumetric imaging for ultrahigh-speed brain mapping at synaptic resolution

Molecular transport through primary human small intestinal monolayers by culture on a collagen scaffold with a gradient of chemical cross-linking

Comparison of the Detection Rates of Different Diagnostic Methods for Primary Peripheral Lung Cancer

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