MRI-R2: Development of an long wavelength nonlinear optical microscope for harmonic and autofluorescence imaging of biological tissues

MRI-R2：开发用于生物组织谐波和自发荧光成像的长波长非线性光学显微镜

• 批准号: 0959525
• 负责人: Paul Campagnola
• 金额: $ 43.5万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2013-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0959525&HistoricalAwards=false
• 关键词: MRI; R2; Development; long; wavelength

0959525Campagnola"This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)."Problems of overlapping spectra, photobleaching, low signal to noise and light scatter pose serious limitations on what can be achieved in practice in imaging tissues by fluorescence. Over the last 10-15 years nonlinear optical microscopy techniques have been developed and adopted widely, largely due to their ability to overcome many of these limitations. Most current nonlinear optical microscopes use excitation wavelengths between 700-1000 nm, as this is the standard tuning range of the widely used Ti:Sapphire femtosecond lasers. While these near infrared (NIR) wavelengths provide significantly deeper penetration (few hundred microns vs. 10 s of microns) into tissue than achievable with confocal laser scanning microscopes using visible excitation sources (e.g. 488, 514 nm), it would be of great biological and biomedical benefit to further improve upon this limit to achieve about 1 mm for imaging the collagen structure associated with tumors and other pathologies. To address this need, this proposal will engage researchers from biomedical engineering, physics, plant biology, cancer biology, and biophotonics to build a nonlinear microscopy platform capable of exciting intrinsic and extrinsic fluorophores from 680nm to 1300 nm. The instrument will be constructed from a Ti:Sapphire oscillator/ optical parametric oscillator (OPO) and a homebuilt laser scanning microscope optimized for longer wavelength excitation. When complete, the instrument will afford detection of 2 and 3-photon excited fluorescence, Second Harmonic Generation and Third Harmonic Generation. We anticipate that using the 1000-1300 nm tuning range provided by the optical parametric oscillator will provide an additional 2-3 fold increase in imaging depth in tissues. The successful construction and implementation of the instrumentation will enable a wide range of dynamic cell studies ranging from plant and cell models to animal models of breast cancer and ovarian cancer. These studies will thoroughly examine the potential of long wavelengths for deeper imaging and improved excitation of fluorophores that naturally excite further into the NIR. This construction will lay down the experimental framework for future biological studies that may benefit from longer wavelength based nonlinear optical approaches.

0959525 Campagnola“该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。“重叠光谱、光漂白、低信噪比和光散射等问题严重限制了荧光组织成像的实际应用。在过去的10-15年中，非线性光学显微镜技术得到了广泛的发展和采用，主要是由于它们能够克服许多这些限制。目前大多数非线性光学显微镜使用700-1000 nm之间的激发波长，因为这是广泛使用的Ti：Sapphire飞秒激光器的标准调谐范围。虽然这些近红外（NIR）波长提供显著更深的穿透（几百微米与10微米）进入组织比使用可见激发源的共焦激光扫描显微镜可实现的效果（例如488、514 nm），进一步改善这一限制以实现约1 mm的肿瘤相关胶原结构成像将具有巨大的生物学和生物医学益处和其他病理学。为了满足这一需求，该提案将吸引生物医学工程、物理学、植物生物学、癌症生物学和生物光子学的研究人员来构建一个能够激发680 nm至1300 nm范围内的内在和外在荧光团的非线性显微镜平台。该仪器将由钛：蓝宝石振荡器/光学参量振荡器（OPO）和一个自制的激光扫描显微镜优化更长的波长激发。完成后，该仪器将提供2和3光子激发荧光，二次谐波产生和三次谐波产生的检测。我们预计，使用光学参量振荡器提供的1000-1300 nm调谐范围将使组织中的成像深度增加2-3倍。该仪器的成功构建和实施将使从植物和细胞模型到乳腺癌和卵巢癌动物模型的广泛动态细胞研究成为可能。这些研究将彻底检查长波长的潜力，用于更深的成像和改进荧光团的激发，这些荧光团自然地激发进近红外。这种结构将为未来的生物学研究奠定实验框架，这些研究可能受益于基于更长波长的非线性光学方法。