IDBR: In vivo multifunctional photothermal cytometry

两种方法在生物学研究中占主导地位：亚细胞水平的实验室实验和活生物体中细胞群体的实验。这给我们对生物现象的理解留下了一个持续和反复出现的空白。在试管中充分复制生物体中重要的全部环境因素是不可能的，并且在具有足够分辨率和灵敏度的研究单个细胞的结构和活动方面，全动物研究目前是有限的，特别是在生物流动背景下。该奖项将通过开发一种基于固有振动光谱对比的多参数光热细胞仪来填补这一空白，该仪器可在静态和流动条件下实时研究生物体中的单个细胞，具有分子特异性。该先进的技术平台将采用光谱可调的近红外纳秒脉冲激光器，用于单细胞的非侵入性、快速（毫秒级）检测和成像，具有高光学分辨率、增强的吸收灵敏度（比传统吸收光谱提高4个数量级）和精确的细胞定位。该仪器可用于各种活体动物模型的研究，包括小鼠耳和肠系膜或秀丽隐杆线虫。项目成果涵盖了一个几乎未开发的生物研究领域，该领域涉及使用弱荧光、吸光的细胞微纳米结构作为细胞特异性光热标志物，用于活体中各种生物过程的非侵入性实时研究，如程序性细胞死亡（凋亡）、代谢活动以及环境因素（如氧化应激）对体内平衡、免疫和寿命的影响。长期目标包括增强光热细胞仪在三维成像中的应用，发展光热多光子显微镜，并潜在地整合光热、荧光和光散射技术。这项跨学科研究涉及数学家、物理学家、电气工程师、化学家和生物学家，并将包括开发一个培训计划和新的课程，内容涉及先进的光学成像、动物模型和体内细胞研究，以使阿肯色大学医学院和阿肯色大学费耶特维尔分校小石城分校的本科生、研究生和研究生的教育受益。来自派恩布拉夫大学的不同种族的学生和残疾人（通过阿肯色聋人学校）将得到指导。

Two methodologies have dominated in biological studies: Laboratory experiments at the subcellular levels, and experiments examining populations of cells in living organisms. This leaves a persistent and recurrent gap in our understanding of biological phenomena. It is not possible to adequately replicate in a test tube the full repertoire of environmental factors that are important in living organisms, and whole-animal studies currently are limited with regards to studying the structure and activities of individual cells with sufficient resolution and sensitivity, especially in a bioflow context. This award will fill this gap by developing a multiparameter photothermal cytometer for real-time study of individual cells in living organisms, in both static and flow conditions with molecular specificity, based upon intrinsic vibrational spectroscopic contrast. The advanced technical platform with a spectrally tunable near-infrared nanosecond pulse laser will be built for non-invasive, rapid (millisecond scale) detection and imaging of single cells with high optical resolution, enhanced absorption sensitivity (four orders better than in convention absorption spectroscopy), and accurate cell positioning. The instrument can be applicable to studies of various living animal models, including mouse ear and mesentery or C. elegans. The project outcomes encompass an almost unexplored area of biological research associated with the use of weakly fluorescent, light-absorbing cellular micro- and nanostructures as intrinsic cell-specific photothermal markers for the non-invasive real-time study in living organisms of various biological processes such as programmed cell death (apoptosis), metabolic activity, and the influence of environmental factors (e.g., oxidative stress) on homeostasis, immunity, and longevity. Long-term goals include enhancing the photothermal cytometer for use in three-dimensional imaging, developing photothermal multiphoton microscopy, and potentially integrating photothermal, fluorescence, and light scattering techniques. This interdisciplinary research involves mathematicians, physicists, electrical engineers, chemists and biologists, and will include developing a training program and new coursework on advanced optical imaging, animal models, and cell study in vivo to benefit the education of undergraduate, graduate, and postgraduate students at University of Arkansas for Medical Sciences and University of Arkansas at Fayetteville at Little Rock. Students from different ethnic groups from University at Pine Bluff, and disabilities (through Arkansas School for the Deaf) will be mentored.