All-Optical Histology for 3-D Cortical Reconstructions

用于 3D 皮质重建的全光学组织学

• 批准号: 7011226
• 负责人: David Kleinfeld
• 金额: $ 15.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7011226
• 关键词: bioimaging /biomedical imaging; blood vessels; brain circulation; brain imaging /visualization /scanning; brain mapping; capillary; cell nucleus; genetically modified animals; histology; laboratory mouse; method development; neuroanatomy; optical tomography; parietal lobe /cortex; three dimensional imaging /topography

DESCRIPTION (provided by applicant): The advent of ultrashort laser light pulses as laboratory tool has opened up new opportunities to probe and manipulate anatomy and function in nervous systems. Ultrashort pulses are the essential means to drive the nonlinear absorption of light by biomolecules, which leads to a localized region of excitation and forms the basis of two-photon scanning microscopy. More recently, nonlinear optical absorption has been exploited as a means to reliably and reproducibly create micron-sized ablations in brain tissue with a minimum of collateral and thermal damage. These ablations can be use as the driving technology in an all-optical histology, which allows anatomy to be imaged with micrometer resolution throughout the entire brain. These ablations may also be used to perturb neocortical blood flow as a means to probe normal and diseased tissues. Yet much additional effort is required to use and advance the mixture of multi-photon ablation and imaging techniques as a means to enable studies of neuronal and vascular architectonics. Our proposed research concerns the confluence of nonlinear optics and anatomy. . Advance the mixture of multi-photon ablation and imaging to establish all-optical based histology as a standard anatomical tool. This includes the optimization of parameters and the advancement of software for combined ablation and imaging. . Reconstruct cell soma and vasculature positions throughout the vibrissa sensory areas in rat cortex. This information will be used to evaluate essential architectonic parameters, including correlations among cell densities in lateral as well as radial directions, as well as essential metabolic parameters, such as the interconnectivity within the vasculature and the distribution of somata relative to capillaries. These two goals, one technical and the other scientific, are intrinsically linked and will proceed in parallel. The proposed advancements will provide a novel tool for the automation of histology, which underlies an understanding of brain function. We will make this tool readily available to the biomedical community. The proposed model system may substantially improve upon our understanding of the large-scale structure of brain architectonics.

描述（由申请人提供）：超短激光脉冲作为实验室工具的出现为探测和操纵神经系统的解剖结构和功能开辟了新的机会。超短脉冲是驱动生物分子对光的非线性吸收的基本手段，从而产生局部激发区域，并构成双光子扫描显微镜的基础。最近，非线性光学吸收已被用作一种可靠且可重复地在脑组织中产生微米级消融的手段，同时将附带和热损伤降至最低。这些消融可以用作全光学组织学中的驱动技术，从而可以在整个大脑中以微米分辨率对解剖结构进行成像。这些消融也可用于扰乱新皮质血流，作为探测正常和患病组织的手段。然而，需要付出更多的努力来使用和推进多光子消融和成像技术的结合，作为研究神经元和血管结构的手段。 我们提出的研究涉及非线性光学和解剖学的融合。 。推进多光子消融和成像的结合，以建立基于全光学的组织学作为标准解剖工具。这包括参数的优化以及用于组合消融和成像的软件的进步。 。重建大鼠皮质整个触须感觉区域的细胞体和脉管系统位置。该信息将用于评估基本的结构参数，包括横向和径向细胞密度之间的相关性，以及基本的代谢参数，例如脉管系统内的互连性和体细胞相对于毛细血管的分布。 这两个目标，一个是技术目标，另一个是科学目标，有着内在的联系，并将并行进行。 所提出的进展将为组织学自动化提供一种新颖的工具，这是理解大脑功能的基础。我们将使生物医学界可以轻松使用该工具。所提出的模型系统可能会大大提高我们对大脑构造学大规模结构的理解。

项目成果

Vectorization of optically sectioned brain microvasculature: learning aids completion of vascular graphs by connecting gaps and deleting open-ended segments.

• DOI: 10.1016/j.media.2012.06.004
• 发表时间: 2012-08
• 期刊: MEDICAL IMAGE ANALYSIS
• 影响因子: 10.9
• 作者: Kaufhold, John P.;Tsai, Philbert S.;Blinder, Pablo;Kleinfeld, David
• 通讯作者: Kleinfeld, David

All-optical in situ histology of brain tissue with femtosecond laser pulses.

使用飞秒激光脉冲对脑组织进行全光学原位组织学。

• DOI: 10.1101/pdb.prot073858
• 发表时间: 2013
• 期刊: Cold Spring Harbor protocols
• 作者: Tsai,PhilbertS;Blinder,Pablo;Squier,JeffreyA;Kleinfeld,David
• 通讯作者: Kleinfeld,David

The cortical angiome: an interconnected vascular network with noncolumnar patterns of blood flow.

• DOI: 10.1038/nn.3426
• 发表时间: 2013-07
• 期刊: NATURE NEUROSCIENCE
• 影响因子: 25
• 作者: Blinder, Pablo;Tsai, Philbert S.;Kaufhold, John P.;Knutsen, Per M.;Suhl, Harry;Kleinfeld, David
• 通讯作者: Kleinfeld, David