Microsecond Raman spectroscopy: Assessing single cell metabolism in a vital organism

微秒拉曼光谱:评估重要生物体中的单细胞代谢

基本信息

  • 批准号:
    8934127
  • 负责人:
  • 金额:
    $ 23.72万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2014
  • 资助国家:
    美国
  • 起止时间:
    2014-09-30 至 2017-08-31
  • 项目状态:
    已结题

项目摘要

 DESCRIPTION: Quantitation of metabolic conversions in live cells and in real time is essential to determining how a cell responds to an intervention such as drug treatment or exposure to a risk factor. Nevertheless, most of our knowledge about cellular content is derived from in vitro analysis of isolated cells or measurement of tissue homogenates, either by biochemical assays, omics or sequencing technologies. This gap highlights a need of developing new techniques that are able to repetitively assess the same single cell in a vital organism. We propose to develop a new Raman imaging platform to enable repetitive assessment of single cell metabolism in a vital organism, using C.elegans as a test bed. Our innovation is spectral scanning of a femtosecond pulse at the Fourier plane of an angle-to wavelength pulse shaper, through which a SRS spectrum can be acquired on the scale of 20 ms per pixel. The long-term goal of our research is developing next generation technology to enable quantitative analysis of single live cell response to a stimuli or a treatment in 3D cultures or live animals. The specific objectives of this R21 application are constructing a ms time scale spectroscopic imaging system and longitudinally assessing the fat metabolism in vital C.elegans. An interdisciplinary team has been formed. Dr. Ji-Xin Cheng (PI) is an expert in label-free spectroscopic imaging. Dr. Heidi Tissenbaum (co-PI) is an expert in dissecting molecular mechanisms of the aging process using C.elegans as a model organism. The two investigators have an established collaboration in developing coherent Raman scattering microscopy to study lipid metabolism in live C.elegans. In feasibility studies, the team has demonstrated hyperspectral SRS imaging of lipid oxidation, lipid desaturation, and cholesterol storage in adult worms using fingerprint Raman bands. Moreover, the Cheng lab recently demonstrated spectral modulation SRS imaging with an angle-to- wavelength pulse shaper. Such development paves the foundation for acquisition of a SRS spectrum on the ms time scale. Our hypothesis that ms Raman spectroscopy is able to longitudinally assess lipid metabolism in vital C.elegans during aging, diet restriction or overfeeding. To test this hypothesis, we will design and construct, and test a microsecond Raman spectral imaging with an angle to wavelength pulse shaper. We will then use the system to longitudinally assess energy metabolism of single cells in wild-type and mutant C. elegans. Though C.elegans is used as a test bed, our platform heralds a broader impact on biomedical research via assessing single live cell response to an intervention, including monitoring live cell response to a risk factor or tissue regeneration in response to a stimulus.
 产品说明:活细胞中代谢转化的定量和真实的时间对于确定细胞如何响应干预(例如药物治疗或暴露于风险因素)是必不可少的。然而,我们对细胞内容物的大部分了解都来自于对分离细胞的体外分析或对组织匀浆的测量,无论是通过生物化学测定、组学还是测序技术。这一差距凸显了开发新技术的必要性,这些技术能够重复评估重要生物体中的相同单细胞。我们建议开发一种新的拉曼成像平台,使重复评估的单细胞代谢在一个重要的有机体,使用秀丽隐杆线虫作为测试床。我们的创新是光谱扫描的飞秒脉冲在傅立叶平面的角度波长脉冲整形器,通过它可以获得的SRS光谱的规模为20毫秒每像素。我们研究的长期目标是开发下一代技术,以定量分析单个活细胞对3D培养物或活动物中刺激或治疗的反应。这个R21应用程序的具体目标是构建一个ms时间尺度的光谱成像系统,并纵向评估重要的秀丽隐杆线虫的脂肪代谢。组建了一支跨学科的团队。Ji-Xin Cheng博士(PI)是无标记光谱成像方面的专家。Heidi Tissenbaum博士(合作PI)是使用秀丽隐杆线虫作为模式生物解剖衰老过程分子机制的专家。这两个研究人员已经建立了合作,在发展相干拉曼散射显微镜研究脂质代谢活秀丽隐杆线虫。在可行性研究中,研究小组已经证明了使用指纹拉曼带对蠕虫的脂质氧化、脂质去饱和和胆固醇储存进行高光谱SRS成像。此外,Cheng实验室最近展示了使用角度到波长脉冲整形器的光谱调制SRS成像。这种发展为ms时间尺度上的SRS频谱的获取奠定了基础。我们假设ms拉曼光谱能够纵向评估衰老、饮食限制或过度喂养期间重要秀丽隐杆线虫的脂质代谢。为了验证这一假设,我们将设计、构造并测试一个具有角度波长脉冲整形器的微秒级拉曼光谱成像。然后,我们将使用该系统纵向评估野生型和突变型C中单细胞的能量代谢。优美的虽然秀丽隐杆线虫被用作测试平台,但我们的平台通过评估单个活细胞对干预的反应,包括监测活细胞对风险因素的反应或对刺激的组织再生,预示着对生物医学研究的更广泛影响。

项目成果

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Ji-Xin Cheng其他文献

Ji-Xin Cheng的其他文献

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{{ truncateString('Ji-Xin Cheng', 18)}}的其他基金

2023 Chemical Imaging Gordon Research Conferences
2023 年化学成像戈登研究会议
  • 批准号:
    10605394
  • 财政年份:
    2023
  • 资助金额:
    $ 23.72万
  • 项目类别:
Sub-millimeter precision wireless neuromodulation using a microwave split ring resonator
使用微波开口环谐振器的亚毫米精度无线神经调节
  • 批准号:
    10669784
  • 财政年份:
    2022
  • 资助金额:
    $ 23.72万
  • 项目类别:
High-content High-speed Chemical Imaging of Metabolic Reprogramming by Integration of Advanced Instrumentation and Data Science
通过先进仪器和数据科学的集成进行代谢重编程的高内涵高速化学成像
  • 批准号:
    10543185
  • 财政年份:
    2022
  • 资助金额:
    $ 23.72万
  • 项目类别:
High-content High-speed Chemical Imaging of Metabolic Reprogramming by Integration of Advanced Instrumentation and Data Science
通过先进仪器和数据科学的集成进行代谢重编程的高内涵高速化学成像
  • 批准号:
    10344774
  • 财政年份:
    2022
  • 资助金额:
    $ 23.72万
  • 项目类别:
Sub-millimeter precision wireless neuromodulation using a microwave split ring resonator
使用微波开口环谐振器的亚毫米精度无线神经调节
  • 批准号:
    10516429
  • 财政年份:
    2022
  • 资助金额:
    $ 23.72万
  • 项目类别:
Mapping Cancer Metabolism by Mid-infrared Photothermal Microscopy
通过中红外光热显微镜绘制癌症代谢图
  • 批准号:
    10491322
  • 财政年份:
    2021
  • 资助金额:
    $ 23.72万
  • 项目类别:
Mapping Cancer Metabolism by Mid-infrared Photothermal Microscopy
通过中红外光热显微镜绘制癌症代谢图
  • 批准号:
    10271761
  • 财政年份:
    2021
  • 资助金额:
    $ 23.72万
  • 项目类别:
Mapping Cancer Metabolism by Mid-infrared Photothermal Microscopy
通过中红外光热显微镜绘制癌症代谢图
  • 批准号:
    10675665
  • 财政年份:
    2021
  • 资助金额:
    $ 23.72万
  • 项目类别:
Vibrational Spectroscopic Imaging to Unveil Hidden Signatures in Living Systems
振动光谱成像揭示生命系统中隐藏的特征
  • 批准号:
    10206200
  • 财政年份:
    2020
  • 资助金额:
    $ 23.72万
  • 项目类别:
Vibrational Spectroscopic Imaging to Unveil Hidden Signatures in Living Systems
振动光谱成像揭示生命系统中隐藏的特征
  • 批准号:
    10660979
  • 财政年份:
    2020
  • 资助金额:
    $ 23.72万
  • 项目类别:

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