IDBR: A Focus-Engineered Coherent Anti-Stokes Raman Scattering (FE-CARS) Microscope for Biological Imaging

IDBR：用于生物成像的 Focus 工程相干反斯托克斯拉曼散射 (FE-CARS) 显微镜

• 批准号: 0754624
• 负责人: Eric Potma
• 金额: $ 26.85万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0754624&HistoricalAwards=false
• 关键词: IDBR; Focus; Engineered; Coherent; Anti

This award is for developing improved contrast in Coherent Anti-Stokes Raman Scattering [CARS] Microscopy through Focus-Engineering [FE-CARS]. Coherent anti-Stokes Raman scattering (CARS) microscopy has rapidly ascended as a noninvasive tool for imaging cells and tissues with chemical selectivity. Based on its unique vibrational selectivity to lipids, proteins, and DNA, the full range of potential CARS applications to biological problems is vast. Nonetheless, due to the presence of a ubiquitous nonresonant background and the weak vibrational response of several molecular groups, applications have been limited to imaging of lipids so far. To open up a new range of applications for CARS microscopy in biology and biomedicine, background suppression and contrast improvement methods are imperative. This work introduces a new approach to CARS microscopy, which provides immediate contrast improvement. The approach is based on controlling the spatial interference of the CARS waves emanating from the focal volume. Emission control is achieved by spatial phase shaping, or focal engineering, of the incident laser pulses using a spatial light modulator (SLM). The use of appropriate phase masks allows controlled destructive interference of unwanted background contributions, while constructive interference accentuates vibrationally resonant objects. A focus-engineered (FE-) CARS microscope will be developed, with the following milestones: 1) Characterizing and optimizing contrast improvement with FE-CARS through computer-controlled phase shaping of the incident transverse beam profiles; 2) Combining FE-CARS with two-photon excited fluorescence (TPEF) and focus-engineered second harmonic generation (SE-SHG) for multimodal imaging of tissues and cells. FE-CARS multimodal imaging will be tested on a) lipid depositions and calcium hydroxyapapatite crystals; b) structural changes to collagen fibers under the influence of glycerol or dimethyl-sulfoxide. The proposed microscope development is anticipated to have far reaching implications for biological imaging studies. The proposed instrument will be used for training graduate students, undergraduate students and pre-college students in the field of nonlinear microscopy. The instrument will be used in several outreach programs. Among these programs, the PI organized the Minority Science Program of the School of Biological Sciences at the university and a 4-week Summer School for high school students.

该奖项是为了表彰通过聚焦工程[FE-CARS]在相干反斯托克斯拉曼散射[CARS]显微技术中提高对比度的成果。相干反斯托克斯拉曼散射(CARS)显微镜作为一种对细胞和组织进行化学选择性成像的非侵入性工具而迅速发展起来。基于其对脂质、蛋白质和DNA的独特振动选择性，CARS在生物问题上的全部潜在应用范围是巨大的。尽管如此，由于普遍存在的非共振背景和几个分子基团的微弱振动响应，到目前为止，应用仅限于对脂质的成像。为了开拓CARS显微镜在生物学和生物医学中的新应用范围，背景抑制和对比度改善方法势在必行。这项工作引入了一种新的CARS显微镜方法，它可以立即提高对比度。该方法是基于控制焦体积发出的CARS波的空间干扰。发射控制是通过使用空间光调制器(SLM)对入射激光脉冲进行空间相位整形或聚焦工程来实现的。使用适当的相位掩模允许对不想要的背景成分进行受控的破坏性干涉，而建设性干涉则突出振动共振的对象。将开发聚焦工程(FE-)CARS显微镜，其里程碑如下：1)通过计算机控制的入射横向光束轮廓的相位整形，表征和优化FE-CARS的对比度改善；2)将FE-CARS与双光子激发荧光(TPEF)和聚焦工程二次谐波(SE-SHG)相结合，用于组织和细胞的多模成像。Fe-CARS多模式成像将测试a)脂肪沉积和钙羟基磷灰石晶体；b)在甘油或二甲基亚砜影响下胶原纤维的结构变化。拟议中的显微镜开发预计将对生物成像研究产生深远的影响。该仪器将用于培养研究生、本科生和预科学生在非线性显微镜领域的知识。该仪器将用于几个外展项目。在这些项目中，PI组织了该大学生物科学学院的少数民族科学项目和为期4周的高中生暑期学校。