Wide-field super-resolved DEEP 3-D microscopy

宽视场超分辨深度 3D 显微镜

• 批准号: 1134561
• 负责人: Kelvin Wagner
• 金额: $ 47.49万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1134561&HistoricalAwards=false
• 关键词: Wide; field; super; resolved; DEEP

1134561WagnerThe authors propose to develop several new capabilities of the novel Doppler Encoded Excitation Pattern (DEEP) microscopy technique that they have recently demonstrated for wide-field Biophotonics imaging applications. DEEP microscopy employs traveling-wave structured illumination and a sensitive high-speed single-element detector (instead of an imaging detector array) to sequentially probe the complex spatial frequencies (Fourier coefficients) of the sample using either fluoresced or scattered light. Tomographic algorithms are utilized to synthesize either an extended depth of field (DOF) 2D or, alternatively, with additional Fourier components a full 3D image. The authors have previously demonstrated that by sequentially illuminating a fluorescent sample with traveling frequency-swept sinusoidal intensity fringe patterns of diverse orientations, the DEEP technique can be used to synthesize a 2D image of a ¡­ mm3 volume that resolves diffraction limited wavelength-scale features, all in focus simultaneously, thereby providing a 1000-fold increase in DOF over a conventional microscope. Three new directions enhancing DEEP microscopy for wide-field biophotonic imaging applications will be pursued: high-speed 3D tomographic imaging multiplexed patterns created with crossed acoustooptic Bragg cells; using illumination coherence gating to extend the depth of high-resolution fluorescence imaging in strongly scattering biological tissue by utilizing broadband light; and super-resolution DEEP imaging microscopy exploiting fluorescence saturation. Each of these research directions will demonstrate a novel wide-field imaging modality for studying biological processes faster, deeper, and smaller than possible with existing techniques.

1134561Wagner作者建议开发几种新的多普勒编码激发模式(Depth)显微技术，他们最近展示了这种技术在广域生物光子学成像中的应用。深度显微镜采用行波结构照明和灵敏的高速单元件探测器(而不是成像探测器阵列)，使用荧光或散射光顺序探测样品的复杂空间频率(傅里叶系数)。层析算法被用来合成扩展景深(DOF)2D，或者可替换地，利用附加的傅立叶分量来合成全3D图像。作者先前已经证明，通过顺序照明具有不同方向的行频扫描正弦强度条纹图案的荧光样品，深度技术可以用于合成体积为？mm3的2D图像，该图像可以同时分辨衍射有限的波长尺度特征，从而提供比传统显微镜1000倍的自由度。三个新的方向将被用于增强广域生物光子成像的深度显微镜：高速3D层析成像多路图案由交叉声光Bragg池创建；利用照明相干门利用宽带光在强散射生物组织中扩展高分辨率荧光成像的深度；以及利用荧光饱和的超分辨率深度成像显微镜。这些研究方向中的每一个都将展示一种新的广域成像模式，用于研究生物过程，比现有技术更快、更深、更小。