Densely Multiplexed Fluorescence Imaging by Fourier Transform Fluorometry

通过傅里叶变换荧光测定法进行密集多重荧光成像

• 批准号: 7513222
• 负责人: Leilei PENG
• 金额: $ 9万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7513222
• 关键词: Award; Biological Markers; Biomedical Research; Cells; Color; Compatible; Confocal Microscopy; Data; Development; Devices; Diagnosis; Disease; Fluorescence; Fluorescence Microscopy; Fluorometry; Foundations; Fourier Transform; Fourier transform spectroscopy; Frequencies; Future; Goals; Guidelines; Image; In Vitro; Individual; Label; Laboratories; Life; Maps; Measurable; Measurement; Measures; Mentors; Mentorship; Methods; Microscope; Microscopic; Microscopy; Modification; Molecular; Mus; Numbers; Optical Methods; Optics; Performance; Phase; Photobiology; Photochemistry; Photochemotherapy; Property; Range; Research; Research Personnel; Resolution; Resources; Running; Sampling; Scientist; Series; Speed; Standards of Weights and Measures; System; Techniques; Technology; Testing; Time; Training; base; bioimaging; career; cellular imaging; computerized data processing; data acquisition; density; emission spectroscopy; fluorescence imaging; fluorophore; frontier; improved; instrument; macrophage; medical specialties; millisecond; nanosecond; new technology; next generation; novel; programs; reconstruction; research study; response

DESCRIPTION (provided by applicant): The goal of this K99/ROO proposal is to develop a new platform for highly multiplexed fluorescence microscopy, which could allow tens to hundreds of biomarkers to be visualized in living cells. At present, multiple florescent markers are identified using differences in fluorescence emission wavelength. Here we propose to increase multiplexing density by simultaneously detecting different fluorescent markers with unique excitation, emission, and lifetime properties. The device that makes this possible is a novel Fourier transform fluorometer that is capable of measuring these fluorescence signatures in parallel. This technology has been pioneered by Dr. Leilei Peng, who is currently a Research Fellow at the Wellman Center for Photomedicine. Currently, the Fourier fluorometer is only capable of making single point measurements over a limited range of fluorescence wavelengths and lifetimes. For this proposal, Dr. Peng will develop this technology further to dramatically expand the range of measurable wavelengths and lifetimes. Data acquisition speeds will also be increased to make this method compatible with microscopic imaging. Following these modifications, the fluorometer will be incorporated into a confocal microscopy imaging system and dense multiplexing will be tested and validated. The research will culminate with a demonstration of multiplexed microscopy in vitro. Guillermo J. Tearney, who runs a laboratory that develops microscopic optical methods for disease diagnosis, will mentor Dr. Peng and supervise development of the spectrometer and microscopy system. Co-mentor Tayyaba Hasan is a renowned scientist in photochemistry and photobiology with specialties in photodynamic therapy and fluorescence imaging. Dr. Hasan will assist with development and selection of fluorescent labels with distinct excitation, emission, and lifetime properties as well as functionalization of these labels for experiments in living cells. The opportunities provided by this award will not only allow Dr. Peng to develop this novel optical technique for biomedical imaging, but will also provide her with valuable mentorship, training, and resources to launch her career as an independent academic researcher.

描述（由申请人提供）：该K99/ROO提案的目标是开发一种用于高度多重荧光显微镜的新平台，该平台可以使活细胞中的数十至数百种生物标志物可视化。目前，使用荧光发射波长的差异来鉴定多种荧光标记物。在这里，我们建议通过同时检测具有独特的激发，发射和寿命特性的不同荧光标记来增加复用密度。使这成为可能的设备是一种新型的傅里叶变换荧光计，能够并行测量这些荧光特征。这项技术是由Leilei Peng博士开创的，他目前是Wellman光医学中心的研究员。目前，傅立叶荧光计只能在有限的荧光波长和寿命范围内进行单点测量。对于该提案，彭博士将进一步开发该技术，以大幅扩展可测量波长和寿命的范围。数据采集速度也将提高，使这种方法与显微成像兼容。这些修改后，荧光计将被纳入共聚焦显微镜成像系统和密集的多路复用将进行测试和验证。这项研究将以体外多重显微镜的演示而告终。Guillermo J. Tearney经营着一个开发用于疾病诊断的显微光学方法的实验室，他将指导彭博士并监督光谱仪和显微镜系统的开发。共同导师Tayyaba Hasan是光化学和光生物学领域的著名科学家，擅长光动力疗法和荧光成像。Hasan博士将协助开发和选择具有不同激发，发射和寿命特性的荧光标记，以及这些标记的功能化，用于活细胞实验。该奖项提供的机会不仅将使彭博士能够开发这种用于生物医学成像的新颖光学技术，还将为她提供宝贵的指导，培训和资源，以启动她作为独立学术研究人员的职业生涯。