scholarly journals Multi-immersion open-top light-sheet microscope for high-throughput imaging of cleared tissues

2019 ◽  
Author(s):  
Adam K. Glaser ◽  
Nicholas P. Reder ◽  
Ye Chen ◽  
Chengbo Yin ◽  
Linpeng Wei ◽  
...  
Keyword(s):  
High Throughput ◽  
Light Sheet ◽  
Optical Clearing ◽  
Clinical Specimens ◽  
Light Sheet Microscopy ◽  
Clinical Laboratories ◽  
Recent Advances ◽  
Large Numbers ◽  
Molecular Information ◽  
High Throughput Imaging

AbstractRecent advances in optical clearing and light-sheet microscopy have provided unprecedented access to structural and molecular information from intact tissues. However, current light-sheet microscopes have imposed constraints on the size, shape, number of specimens, and compatibility with various clearing protocols. Here we present a multi-immersion open-top light-sheet microscope that enables simple mounting of multiple specimens processed with a variety of protocols, which will facilitate wider adoption by preclinical researchers and clinical laboratories.One Sentence SummaryGlaseret al.describe a multi-immersion open-top light-sheet microscope that enables simple and high-throughput imaging of large numbers of preclinical and clinical specimens prepared with a variety of clearing protocols.

scholarly journals High-throughput live imaging using Light Sheet Microscopy

2020 ◽  
Author(s):  
Emilio J. Gualda ◽  
Matteo Bernardello ◽  
Maria Marsal ◽  
Pablo Loza Alvarez
Keyword(s):  
High Throughput ◽  
Light Sheet ◽  
Live Imaging ◽  
Light Sheet Microscopy

scholarly journals Quantitative neuroanatomy of all Purkinje cells with light sheet microscopy and high-throughput image analysis

2015 ◽  
Vol 9 ◽  
Author(s):  
Ludovico Silvestri ◽  
Marco Paciscopi ◽  
Paolo Soda ◽  
Filippo Biamonte ◽  
Giulio Iannello ◽  
...  
Keyword(s):  
Image Analysis ◽  
Purkinje Cells ◽  
High Throughput ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals A hybrid open-top light-sheet microscope for multi-scale imaging of cleared tissues

2021 ◽  
Author(s):  
Adam Glaser ◽  
Kevin Bishop ◽  
Lindsey Barner ◽  
Etsuo Susaki ◽  
Shimpei Kubota ◽  
...  
Keyword(s):  
Hybrid System ◽  
High Throughput ◽  
Light Sheet ◽  
Refractive Indices ◽  
Volumetric Imaging ◽  
Tissue Clearing ◽  
Sample Geometry ◽  
Multi Scale ◽  
Light Sheet Microscopy ◽  
User Friendly

Abstract Light-sheet microscopy has emerged as the preferred means for high-throughput volumetric imaging of cleared tissues. However, there is a need for a user-friendly system that can address imaging applications with varied requirements in terms of resolution (mesoscopic to sub-micrometer), sample geometry (size, shape, and number), and compatibility with tissue-clearing protocols and sample holders of various refractive indices. We present a ‘hybrid’ system that combines a novel non-orthogonal dual-objective and conventional (orthogonal) open-top light-sheet architecture for versatile multi-scale volumetric imaging.

scholarly journals Subvoxel light-sheet microscopy for high-resolution high-throughput volumetric imaging of large biomedical specimens

2019 ◽  
Vol 1 (01) ◽  
pp. 1 ◽  
Author(s):  
Peng Fei ◽  
Jun Nie ◽  
Juhyun Lee ◽  
Yichen Ding ◽  
Shuoran Li ◽  
...  
Keyword(s):  
High Resolution ◽  
High Throughput ◽  
Light Sheet ◽  
Volumetric Imaging ◽  
Light Sheet Microscopy

scholarly journals Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains

2015 ◽  
Vol 2 (4) ◽  
pp. 041404 ◽  
Author(s):  
M. Caroline Müllenbroich ◽  
Ludovico Silvestri ◽  
Leonardo Onofri ◽  
Irene Costantini ◽  
Marcel van’t Hoff ◽  
...  
Keyword(s):  
Data Management ◽  
High Throughput ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals Sub-voxel light-sheet microscopy for high-resolution, high-throughput volumetric imaging of large biomedical specimens

2018 ◽  
Author(s):  
Peng Fei ◽  
Jun Nie ◽  
Juhyun Lee ◽  
Yichen Ding ◽  
Shuoran Li ◽  
...  
Keyword(s):  
High Resolution ◽  
High Throughput ◽  
Light Sheet ◽  
Large Field ◽  
Mouse Heart ◽  
Processing Unit ◽  
Intact Mouse ◽  
Volumetric Imaging ◽  
Light Sheet Microscopy ◽  
Wide Range

A key challenge when imaging whole biomedical specimens is how to quickly obtain massive cellular information over a large field of view (FOV). Here, we report a sub-voxel light-sheet microscopy (SLSM) method enabling high-throughput volumetric imaging of mesoscale specimens at cellular-resolution. A non-axial, continuous scanning strategy is used to rapidly acquire a stack of large-FOV images with three-dimensional (3-D) nanoscale shifts encoded. Then by adopting a sub-voxel-resolving procedure, the SLSM method models these low-resolution, cross-correlated images in the spatial domain and iteratively recovers a 3-D image with improved resolution throughout the sample. This technique can surpass the optical limit of a conventional light-sheet microscope by more than three times, with high acquisition speeds of gigavoxels per minute. As demonstrated by quick reconstruction (minutes to hours) of various samples, e.g., 3-D cultured cells, an intact mouse heart, mouse brain, and live zebrafish embryo, the SLSM method presents a high-throughput way to circumvent the tradeoff between intoto mapping of large-scale tissue (>100 mm3) and isotropic imaging of single-cell (~1-μm resolution). It also eliminates the need of complicated mechanical stitching or precisely modulated illumination, using a simple light-sheet setup and fast graphics-processing-unit (GPU)-based computation to achieve high-throughput, high-resolution 3-D microscopy, which could be tailored for a wide range of biomedical applications in pathology, histology, neuroscience, etc.

scholarly journals An orthotopic glioblastoma animal model suitable for high-throughput screenings

2018 ◽  
Vol 20 (11) ◽  
pp. 1475-1484 ◽  
Author(s):  
Linda Pudelko ◽  
Steven Edwards ◽  
Mirela Balan ◽  
Daniel Nyqvist ◽  
Jonathan Al-Saadi ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
Large Scale ◽  
Light Sheet ◽  
Time Lapse ◽  
Zebrafish Model ◽  
Light Sheet Microscopy ◽  
Disease Biology ◽  
Vertebrate Model

Abstract Background Glioblastoma (GBM) is an aggressive form of brain cancer with poor prognosis. Although murine animal models have given valuable insights into the GBM disease biology, they cannot be used in high-throughput screens to identify and profile novel therapies. The only vertebrate model suitable for large-scale screens, the zebrafish, has proven to faithfully recapitulate biology and pathology of human malignancies, and clinically relevant orthotopic zebrafish models have been developed. However, currently available GBM orthotopic zebrafish models do not support high-throughput drug discovery screens. Methods We transplanted both GBM cell lines as well as patient-derived material into zebrafish blastulas. We followed the behavior of the transplants with time-lapse microscopy and real-time in vivo light-sheet microscopy. Results We found that GBM material transplanted into zebrafish blastomeres robustly migrated into the developing nervous system, establishing an orthotopic intracranial tumor already 24 hours after transplantation. Detailed analysis revealed that our model faithfully recapitulates the human disease. Conclusion We have developed a robust, fast, and automatable transplantation assay to establish orthotopic GBM tumors in zebrafish. In contrast to currently available orthotopic zebrafish models, our approach does not require technically challenging intracranial transplantation of single embryos. Our improved zebrafish model enables transplantation of thousands of embryos per hour, thus providing an orthotopic vertebrate GBM model for direct application in drug discovery screens.

scholarly journals Automated screening by 3D light-sheet microscopy with high spatial and temporal resolution reveals mitotic phenotypes

2020 ◽  
Author(s):  
Björn Eismann ◽  
Teresa G Krieger ◽  
Jürgen Beneke ◽  
Ruben Bulkescher ◽  
Lukas Adam ◽  
...  
Keyword(s):  
Cell Cultures ◽  
High Throughput ◽  
Light Exposure ◽  
Epigenetic Modification ◽  
Fluorescent Microscopy ◽  
Light Sheet ◽  
Drug Candidate ◽  
Light Sheet Microscopy ◽  
Phenotype Classification ◽  
3D Cell Cultures

Abstract3D cell cultures enable the in vitro study of dynamic biological processes such as the cell cycle, but their use in high-throughput screens remains impractical with conventional fluorescent microscopy. Here, we present a screening workflow for the automated evaluation of mitotic phenotypes in 3D cell cultures by light-sheet microscopy. After sample preparation by a liquid handling robot, three-dimensional cell spheroids are imaged for 24 hours in toto with a dual inverted selective plane illumination (diSPIM) microscope with a much improved signal-to-noise ratio, higher imaging speed, isotropic resolution and reduced light exposure compared to a spinning disc confocal microscope. A dedicated high-content image processing pipeline implements convolutional neural network based phenotype classification. We illustrate the potential of our approach by siRNA knock-down and epigenetic modification of 28 mitotic target genes for assessing their phenotypic role in mitosis. By rendering light-sheet microscopy operational for high-throughput screening applications, this workflow enables target gene characterization or drug candidate evaluation in tissue-like 3D cell culture models.

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