Adaptive Tracking and Quantum Imaging for Protein-Protein Interactions
蛋白质-蛋白质相互作用的自适应跟踪和量子成像
基本信息
- 批准号:10706952
- 负责人:
- 金额:$ 49.39万
- 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:2022-09-21 至 2026-08-31
- 项目状态:未结题
- 来源:
- 关键词:AlgorithmsAreaBayesian AnalysisBayesian MethodBindingBiomedical ResearchBlinkingCell membraneCell physiologyCellsCollaborationsColorComputer softwareDataData AnalysesDetectionDevelopmentDiffuseDiffusionDiseaseDyesFeedbackGoalsHypersensitivityImageKineticsLabelLibrariesLightLightingMalignant NeoplasmsMarkov ChainsMeasurementMeasuresMembrane ProteinsMethodsMicroscopeMicroscopyNoiseOpticsPaperParameter EstimationPatternPhasePhotobleachingPhotonsProcessProteinsPublishingPythonsQuantum DotsRecording of previous eventsReportingResearch PersonnelResolutionSamplingSchemeSignal TransductionSourceSpeedTechniquesTechnologyTimeToxic effectUncertaintyVisualizationactive controlbiological systemsdensitydimerfluorophoreheterodyningimaging approachimaging modalityimaging systemimprovedinnovationmolecular imagingmovienovelparticleprotein protein interactionquantumsingle moleculetechnology developmentultra high resolution
项目摘要
Project Summary
This project describes a technology development effort that will generate three complementary methods for
measuring Protein-Protein Interaction (PPI) kinetic rates between membrane proteins. In Aim 1, we develop a
new algorithmic approach for single-particle tracking analysis that generates trajectories from image/movie
data. It is based on Bayesian inference strategy. We then extend this to extracting kinetic rates of PPIs from
two color single particle tracking data. In Aim 2, we develop a ‘smart’ microscope that is capable of adapting
illumination and frame rates when a PPI is imminent (particles diffuse close to each other), thereby reducing
photobleaching, allowing long-term tracking with organic fluorophores and gaining a factor of 10 to 50 in
imaging speed during an interaction. In Aim 3 we implement and demonstrate a ‘quantum imaging’ approach
for making a quantum optimal estimation of the distance between two fluorophores of the same type. This
method can estimate the distance between incoherent point sources with an uncertainty in the distance
measurement near the limit of any possible measurement and substantially better than classical
image-then-estimate approaches. In practice, it gives the benefits of two-color tracking using a simplified
one-color labeling scheme.
Direct detection of PPIs at the single molecule level in living cells is difficult because labeling must be done at
densities low enough for single fluorescent labels to be identified and detected with diffraction limited optics.
Since molecules must come into contact with each other to initiate a PPI, often through diffusion limited
processes, the observed interactions can be temporally rare. The new methods proposed here will allow for
other fluorophores, including organic dyes and fluorescent proteins, to be used to quantify PPIs. Our long-term
goal is to provide a set of methods that can report on the dynamics of various types of PPIs. The rationale for
the proposed Aims is that by providing these new methods, a wider range of PPIs can be studied on living
cells, opening new doors for biomedical research.
This technology development project is a collaboration between three investigators with complementary
expertise. K. Lidke (PI) is an expert on single molecule imaging and microscopy development. F. Becerra
(co-PI) is a world leader in the area of ultra-sensitive measurements of coherent states of light using optimized
photon counting measurements with fast feedback. D. Lidke (co-I) has developed and applied innovative single
molecule techniques to quantify PPI in living cells.
项目摘要
该项目描述了一项技术开发工作,将产生三种互补的方法,
测量膜蛋白之间的蛋白质-蛋白质相互作用(PPI)动力学速率。在目标1中,我们开发了一个
单粒子跟踪分析的新算法方法,从图像/电影生成轨迹
数据它是基于贝叶斯推理策略。然后,我们将其扩展到提取PPI的动力学速率,
双色单粒子跟踪数据。在目标2中,我们开发了一种能够适应
当PPI即将到来时(粒子彼此靠近地扩散),
光漂白,允许有机荧光团的长期跟踪,并获得10至50的因子,
在交互过程中的成像速度。在目标3中,我们实现并演示了一种“量子成像”方法
用于对相同类型的两个荧光团之间的距离进行量子最佳估计。这
该方法可以在距离不确定的情况下估计非相干点源之间的距离
测量接近任何可能的测量极限,并大大优于经典的
先成像后估计方法。在实践中,它使用简化的
单色标签方案。
在活细胞中在单分子水平上直接检测PPI是困难的,因为标记必须在
密度低到足以使单个荧光标记物被衍射限制光学器件识别和检测。
由于分子必须相互接触以启动PPI,通常通过扩散限制
过程中,观察到的相互作用可能是暂时罕见的。这里提出的新方法将允许
其他荧光团,包括有机染料和荧光蛋白,用于定量PPI。我们的长期
目标是提供一套方法,可以报告各种类型的PPI的动态。的理由
提出的目标是,通过提供这些新方法,可以在活体上研究更广泛的PPI
细胞,为生物医学研究打开了新的大门。
该技术开发项目是三名研究人员之间的合作,具有互补性
专业知识K. Lidke(PI)是单分子成像和显微镜开发方面的专家。F. Becerra
(co-PI)是世界领先的超灵敏测量领域的相干态光使用优化的
具有快速反馈的光子计数测量。D. Lidke(co-I)开发并应用了创新的单
分子技术来量化活细胞中的PPI。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Tolerance to aberration and misalignment in a two-point-resolving image inversion interferometer.
两点分辨图像反转干涉仪的像差和失准容差。
- DOI:10.1364/oe.487808
- 发表时间:2023
- 期刊:
- 影响因子:3.8
- 作者:Schodt,DavidJ;Cutler,PatrickJ;Becerra,FranciscoE;Lidke,KeithA
- 通讯作者:Lidke,KeithA
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Francisco Elohim Becerra Chavez其他文献
Francisco Elohim Becerra Chavez的其他文献
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{{ truncateString('Francisco Elohim Becerra Chavez', 18)}}的其他基金
Adaptive Tracking and Quantum Imaging for Protein-Protein Interactions
蛋白质-蛋白质相互作用的自适应跟踪和量子成像
- 批准号:
10296577 - 财政年份:2022
- 资助金额:
$ 49.39万 - 项目类别:
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