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Development of photothermal microscopy for biomedical applications

开发用于生物医学应用的光热显微镜

基本信息

批准号：
8232025
负责人：
Jerome Mertz
金额：
$ 20.46万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8232025
关键词：
Attention Beds Biological Models Blood Cells Cell Culture Techniques Cells Chick Embryo Clinic Communities Confocal Microscopy Contrast Media Cytochromes Deposition Detection Development Diagnosis Environment Erythrocytes FarGo Funding Goals Gold Grant Heating Hela Cells Hemeproteins Hemoglobin Image Lasers Life Light Measurement Measures Microbial Rhodopsins Microscope Microscopy Mitochondria Modeling Molecular Monitor Noise Optics Performance Phase Photons Positioning Attribute Proteins Refractive Indices Research Resolution Rhodopsin Sampling Scanning Screening procedure Signal Transduction Source Specificity Spectrum Analysis Structure System Techniques Technology Testing Thick Three-Dimensional Imaging Time Tissues absorption base bioimaging chromophore density in vivo mutant nanoparticle new technology novel optical imaging public health relevance response success time use tissue phantom two-photon

项目摘要

DESCRIPTION (provided by applicant): The imaging of absorbing (i.e. nonfluorescent) chromophores in thick tissue poses a challenge for microscopists. Recently, a new technique called photothermal microscopy (PM) has been gaining attention, which involves the focusing two laser beams into a sample. One beam (the heating beam) is tuned to a chromophore absorption line while the other is set at wavelength outside any absorption bands (the probe beam). When the heating beam is absorbed, energy is deposited into the tissue, producing a local density fluctuation. This density change results in a small transient change in the refractive index about the chromophore, which is then monitored by the probe beam. In early applications, PM has been shown to track gold nanoparticles in cell cultures with unprecedented sensitivity. More recently, PM has also been shown to be remarkably effective at imaging endogenous chromophores in live cells. Examples include imaging of mitochondria and erythrocytes with 3D spatial resolution comparable to confocal microscopy. As promising as PM is, several open questions still remain. Specifically: is it possible to perform PM in thick tissue, and what exactly are the chromophore species responsible for PM contrast? To date, PM has only been performed with thin samples in a transmitted light configuration. Moreover, the chromophore species responsible for PM contrast are either unknown (in the case of mitochondrial imaging) or speculated (in the case of erythrocyte imaging). We propose to 1) develop a novel scanning PM that can perform photothermal imaging in thick tissue, for the first time, using on one- or two-photon absorption, and 2) unambiguously identify and characterize both existing and new endogenous contrast agents using a photothermal spectroscopy screening platform equipped with an ultra-wide bandwidth (UV to THz) laser. We will initially concentrate on the study of cytochrome in mitochondria, heme protein in blood cells, and channel rhodopsin. A completion of the above aims will be indispensible for PM to gain widespread acceptance in the biomedical imaging community, and will lay the groundwork for a new technology that provides high sensitivity, high resolution absorption contrast with molecular specificity. PUBLIC HEALTH RELEVANCE: We propose to develop an optical microscopy technique that provides ultrahigh sensitivity 3D imaging of absorbing (i.e. non-fluorescent) proteins or molecules in tissue. This technology will be useful for in- vivo imaging research applications and rapid tissue diagnosis in the clinic.

描述（由申请人提供）：厚组织中吸收（即非荧光）发色团的成像对显微镜学家提出了挑战。最近，一种称为光热显微镜 (PM) 的新技术引起了人们的关注，该技术涉及将两束激光聚焦到样品中。将一束（加热束）调谐到发色团吸收线，而另一束设置在任何吸收带之外的波长（探测束）。当热束被吸收时，能量沉积到组织中，产生局部密度波动。这种密度变化导致发色团折射率发生微小的瞬时变化，然后由探测光束监测。在早期应用中，PM 已被证明能够以前所未有的灵敏度追踪细胞培养物中的金纳米粒子。最近，PM 也被证明在活细胞内源生色团成像方面非常有效。例如，以与共焦显微镜相当的 3D 空间分辨率对线粒体和红细胞进行成像。尽管 PM 前景广阔，但仍有几个悬而未决的问题。具体来说：是否可以在厚组织中进行 PM，以及负责 PM 对比度的发色团种类到底是什么？迄今为止，PM 仅在透射光配置中对薄样品进行。此外，负责 PM 对比度的发色团种类要么是未知的（在线粒体成像的情况下），要么是推测的（在红细胞成像的情况下）。我们建议1）开发一种新型扫描PM，首次利用一个或两个光子吸收在厚组织中进行光热成像，2）使用配备超宽带（UV至THz）激光器的光热光谱筛选平台明确识别和表征现有和新的内源性造影剂。我们首先将集中研究线粒体中的细胞色素、血细胞中的血红素蛋白和通道视紫红质。上述目标的完成对于PM在生物医学成像界获得广泛接受是必不可少的，并将为提供高灵敏度、高分辨率吸收对比和分子特异性的新技术奠定基础。公共健康相关性：我们建议开发一种光学显微镜技术，为组织中的吸收（即非荧光）蛋白质或分子提供超高灵敏度 3D 成像。该技术将有助于体内成像研究应用和临床快速组织诊断。