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使用 3D 靶向照明进行视频速率容积神经元成像。

Video-rate volumetric neuronal imaging using 3D targeted illumination.

机构信息

Department of Electrical & Computer Engineering, Boston University, 8 Saint Mary's St., Boston, Massachusetts, 02215, USA.

Department of Biomedical Engineering, Boston University, 44 Cummington Mall, Boston, Massachusetts, 02215, USA.

出版信息

Sci Rep. 2018 May 21;8(1):7921. doi: 10.1038/s41598-018-26240-8.


DOI:10.1038/s41598-018-26240-8
PMID:29784920
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5962542/
Abstract

Fast volumetric microscopy is required to monitor large-scale neural ensembles with high spatio-temporal resolution. Widefield fluorescence microscopy can image large 2D fields of view at high resolution and speed while remaining simple and costeffective. A focal sweep add-on can further extend the capacity of widefield microscopy by enabling extended-depth-of-field (EDOF) imaging, but suffers from an inability to reject out-of-focus fluorescence background. Here, by using a digital micromirror device to target only in-focus sample features, we perform EDOF imaging with greatly enhanced contrast and signal-to-noise ratio, while reducing the light dosage delivered to the sample. Image quality is further improved by the application of a robust deconvolution algorithm. We demonstrate the advantages of our technique for in vivo calcium imaging in the mouse brain.

摘要

快速体积显微镜是监测具有高时空分辨率的大规模神经团的必要手段。宽场荧光显微镜可以以高分辨率和速度成像大的 2D 视场,同时保持简单和具有成本效益。焦点扫描附加组件可以通过实现扩展景深(EDOF)成像进一步扩展宽场显微镜的容量,但存在无法消除离焦荧光背景的问题。在这里,我们通过使用数字微镜设备仅针对聚焦的样本特征,在大大提高对比度和信噪比的同时,减少了施加到样本的光剂量,从而进行 EDOF 成像。通过应用强大的去卷积算法进一步提高了图像质量。我们展示了我们的技术在活体小鼠大脑钙成像中的优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/affc8c951ff6/41598_2018_26240_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/9b5e0aa0fad6/41598_2018_26240_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/32058d425bb1/41598_2018_26240_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/5bc847ad6c48/41598_2018_26240_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/dfcb83a78d72/41598_2018_26240_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/cc7d1afa3f80/41598_2018_26240_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/affc8c951ff6/41598_2018_26240_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/9b5e0aa0fad6/41598_2018_26240_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/32058d425bb1/41598_2018_26240_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/5bc847ad6c48/41598_2018_26240_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/dfcb83a78d72/41598_2018_26240_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/cc7d1afa3f80/41598_2018_26240_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3de/5962542/affc8c951ff6/41598_2018_26240_Fig6_HTML.jpg

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[1]
Video-rate volumetric neuronal imaging using 3D targeted illumination.

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引用本文的文献

[1]
Advancing Multicolor Super-Resolution Volume Imaging: Illuminating Complex Cellular Dynamics.

JACS Au. 2025-6-9

[2]
Compressive streak microscopy for fast sampling of fluorescent reporters of neural activity.

Neurophotonics. 2025-4

[3]
Random Illumination Microscopy: faster, thicker, and aberration-insensitive.

Light Sci Appl. 2025-1-2

[4]
Learning flat optics for extended depth of field microscopy imaging.

Nanophotonics. 2023-8-2

[5]
Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging.

Light Sci Appl. 2024-10-10

[6]
Targeted illumination confocal microscopy enables in vivo voltage imaging in thick tissue.

Nat Methods. 2024-6

[7]
Large-scale deep tissue voltage imaging with targeted-illumination confocal microscopy.

Nat Methods. 2024-6

[8]
Neurophotonics beyond the surface: unmasking the brain's complexity exploiting optical scattering.

Neurophotonics. 2024-9

[9]
DeepDOF-SE: affordable deep-learning microscopy platform for slide-free histology.

Nat Commun. 2024-4-5

[10]
Neurophotonics beyond the Surface: Unmasking the Brain's Complexity Exploiting Optical Scattering.

ArXiv. 2024-3-21

本文引用的文献

[1]
A targeted illumination optical fiber probe for high resolution fluorescence imaging and optical switching.

Sci Rep. 2017-4-3

[2]
Video-rate volumetric functional imaging of the brain at synaptic resolution.

Nat Neurosci. 2017-4

[3]
Extended depth-of-field microscopy with a high-speed deformable mirror.

Opt Lett. 2017-3-1

[4]
Adaptive illumination reduces photobleaching in structured illumination microscopy.

Biomed Opt Express. 2016-9-23

[5]
Fast volumetric calcium imaging across multiple cortical layers using sculpted light.

Nat Methods. 2016-12

[6]
Fast 3D Imaging of Spine, Dendritic, and Neuronal Assemblies in Behaving Animals.

Neuron. 2016-11-23

[7]
Random-access scanning microscopy for 3D imaging in awake behaving animals.

Nat Methods. 2016-12

[8]
Genetically encoded indicators of neuronal activity.

Nat Neurosci. 2016-8-26

[9]
Calcium imaging of neural circuits with extended depth-of-field light-sheet microscopy.

Opt Lett. 2016-3-1

[10]
Simultaneous Denoising, Deconvolution, and Demixing of Calcium Imaging Data.

Neuron. 2016-1-20

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