Open-source miniaturized two-photon microscopes for large field-of-view and volumetric imaging
用于大视场和体积成像的开源小型双光子显微镜
基本信息
- 批准号:10675751
- 负责人:
- 金额:$ 98.22万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-08-15 至 2026-07-31
- 项目状态:未结题
- 来源:
- 关键词:AnimalsAnteriorAxonBehaviorBrainBypassCalciumCellsCollaborationsCommunitiesCoupledCustomCytoplasmic GranulesDecision MakingDendritesEducational process of instructingEducational workshopElementsEnsureExcisionFiberGlutamatesHippocampusImageLateralLettersLinkMemoryMethodsMicroscopeMicroscopyMusNeuronsNeurosciencesOperative Surgical ProceduresOpticsParietal LobePatternPersonsPhotonsPopulationProductionPublishingRattusRecording of previous eventsResearchResearch PersonnelResolutionRoleScanningSocial InteractionSpecific qualifier valueStructureSystemTechniquesTechnologyTestingThalamic structureTissue imagingTissuesWorkcingulate cortexcostdata acquisitiondesigndesign,build,testequipment acquisitionimaging capabilitiesinsightlensmemory encodingmeterminiaturizeneural networknonhuman primatenovelopen sourceopen source toolphotonicssensorspatial memorytheoriestooltwo photon microscopytwo-photonwiki
项目摘要
Abstract:
Single-photon (1P) epifluorescence miniaturized microscopy coupled with genetically encoded calcium sensors
has allowed investigators to record the activity of large populations of identified neurons over days to weeks in
freely behaving animals, answering fundamental questions in neuroscience. Our group's efforts with the UCLA
Miniscope Project have allowed over 600 labs to build and use over 2500 open-source miniaturized microscopes
with expanded capabilities at a small fraction of the cost of those offered by commercial versions, thus
democratizing access. Yet, 1P miniscopes lack the lateral and axial resolution to image activity in fine structures
such as dendrites and axons. In addition, 1P imaging is limited to superficial structures or requires removal of
overlying tissue for imaging of deeper neurons. Two-photon (2P) microscopy has exquisite lateral and axial
resolution and bypasses all of these obstacles. Recent advances in technology have made the construction of
two-photon miniaturized microscopes for mice possible. However, the field of view (FOV) is still limited, and
these microscopes require custom-built optics and cost several hundred thousand dollars to acquire
commercially. We have designed and built a two-photon miniaturized microscope for mice, including a custom-
made objective lens, that allows 2P imaging of an 800 micrometer FOV nearly quadrupling the FOV from the
latest published 2P miniaturized microscope (Mini2P-V1). In this proposal, we will optimize this microscope and
test it in freely behaving mice for axonal, dendritic and deep somatic imaging. This microscope will be tested in
three labs. The Golshani Lab will test the scope with calcium imaging of thalamic axons in anterior cingulate
cortex during social interaction. The Silva Lab will test the scope by performing dendritic calcium and glutamate
imaging in retrosplenial cortex during memory linking. The Shtrahman Lab will test deep imaging capability by
imaging dentate granule neurons through an intact CA1. We will also build a larger miniaturized microscope
suitable for rats and non-human primates with expanded capabilities, including a higher numerical aperture (NA),
large FOV and temporal multiplexing capability to allow volumetric imaging at high frame rates (MiniMux2P).
This microscope will be tested by the Blair Lab to dissect the role of superficial and deep CA1 neurons of rats in
navigation. It will also be tested in the Churchland Lab to image rat posterior parietal cortical neurons during
decision-making tasks. Finally, we will disseminate the technology using our open-source wiki that has already
disseminated miniscope technology to thousands of users. We will provide parts-lists, optical designs and
methods for obtaining custom lens elements. As we have done before, we will educate users through online
videos and hands-on workshops where imaging basics, surgical techniques and analysis tools are demonstrated.
We hope these cutting edge, novel and open-source tools will allow investigators to extend their research beyond
that of what is possible with currently available technology.
摘要:
单光子(1 P)落射荧光微型化显微镜与遗传编码钙传感器耦合
这使得研究人员能够在几天到几周的时间里记录大量已识别神经元的活动,
自由行为的动物,回答神经科学的基本问题。我们小组与加州大学洛杉矶分校的合作
Miniscope项目允许600多个实验室建造和使用2500多个开源微型显微镜
与商业版本相比,其成本仅为一小部分,
民主化访问。然而,1 P微型镜缺乏横向和轴向分辨率,无法在精细结构中成像活动
例如树突和轴突。此外,1 P成像仅限于浅表结构或需要去除
用于更深层神经元的成像。双光子(2 P)显微镜具有精致的横向和轴向
解决方案,并绕过所有这些障碍。最近的技术进步使建造
用于小鼠的双光子微型显微镜成为可能。然而,视场(FOV)仍然有限,并且
这些显微镜需要定制的光学系统,
在商业上。我们已经设计并建造了一个双光子微型显微镜的小鼠,包括定制-
制造的物镜透镜,允许800微米FOV的2 P成像,几乎是FOV的四倍。
最新发布的2 P微型显微镜(Mini2 P-V1)。在这个建议中,我们将优化这个显微镜,
在行为自由的小鼠中测试它的轴突、树突和深部体细胞成像。这台显微镜将在
三个实验室Golshani实验室将通过前扣带回中丘脑轴突的钙成像来测试该范围
在社交互动中。席尔瓦实验室将通过对树突状钙和谷氨酸盐进行测试来测试示波器
在记忆链接时的压后皮质成像。Shtrahman实验室将测试深度成像能力,
通过完整的CA 1成像齿状颗粒神经元。我们还将建造一个更大的微型显微镜
适用于具有扩展能力的大鼠和非人灵长类动物,包括更高的数值孔径(NA),
大FOV和时间多路复用能力,允许以高帧率进行体积成像(MiniMux 2 P)。
这台显微镜将由布莱尔实验室进行测试,以解剖大鼠的浅层和深层CA 1神经元,
导航它也将在Churchland实验室进行测试,以成像大鼠后顶叶皮层神经元,
决策任务。最后,我们将使用我们的开源wiki来传播这项技术,
向成千上万的用户传播微型望远镜技术。我们将提供零件清单,光学设计,
用于获得定制透镜元件的方法。正如我们以前所做的那样,我们将通过在线教育用户,
视频和实践研讨会,展示成像基础知识、手术技术和分析工具。
我们希望这些尖端、新颖和开源的工具能让研究人员将他们的研究扩展到
即现有技术的可能性。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
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Daniel Aharoni其他文献
Daniel Aharoni的其他文献
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{{ truncateString('Daniel Aharoni', 18)}}的其他基金
Open-source miniaturized two-photon microscopes for large field-of-view and volumetric imaging
用于大视场和体积成像的开源小型双光子显微镜
- 批准号:
10516900 - 财政年份:2022
- 资助金额:
$ 98.22万 - 项目类别:
Closed-Loop Systems for Large Scale Spatiotemporal Imaging and Actuation of Neural Activity in Freely Behaving Animals
用于自由行为动物的大规模时空成像和神经活动激活的闭环系统
- 批准号:
10401560 - 财政年份:2022
- 资助金额:
$ 98.22万 - 项目类别:
Closed-Loop Systems for Large Scale Spatiotemporal Imaging and Actuation of Neural Activity in Freely Behaving Animals
用于自由行为动物的大规模时空成像和神经活动激活的闭环系统
- 批准号:
10675440 - 财政年份:2022
- 资助金额:
$ 98.22万 - 项目类别:
Developing long-term neuro-behavioral recording and real-time processing platforms for naturally behaving animals
为自然行为动物开发长期神经行为记录和实时处理平台
- 批准号:
10245927 - 财政年份:2021
- 资助金额:
$ 98.22万 - 项目类别:
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