Chromatic two-photon fluorescence microsopy using band-shifting imaging probes

使用带移成像探针的彩色双光子荧光显微镜

• 批准号: 9456125
• 负责人: Zhiwen Liu
• 金额: $ 22.48万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9456125
• 关键词: Address; Amination; Amines; Biocompatible Materials; Biological Process; Biophotonics; Cancer Detection; Citrates; Citric Acid; Collaborations; Colon Carcinoma; Colorectal Cancer; Detection; Development; Diagnosis; Dyes; Early Diagnosis; Endoscopy; Exhibits; Fluorescence; Fluorescence Microscopy; Fluorescent Probes; Goals; HT29 Cells; Image; Imaging Techniques; Label; Lasers; Lateral; Location; Maps; Mechanics; Medical; Methodology; Methods; Metric System; Microscopy; Modality; Molecular Structure; Molecular Weight; Monitor; Outcome; Penetration; Phototoxicity; Positioning Attribute; Property; Research; Resolution; Scanning; Signal Transduction; Source; Specimen; Speed; Techniques; Tissue imaging; Work; Xenograft Model; Yang; base; biological research; cancer cell; cancer diagnosis; cellular imaging; design; detector; digital; imaging modality; imaging probe; imaging study; imaging system; in vivo; in vivo imaging; innovation; lens; novel; optical fiber; programs; tissue phantom; tool; two-photon

Chromatic two-photon fluorescence microscopy using band-shifting imaging probes Two-photon fluorescence microscopy has emerged as one of the most important imaging modalities for biological research and medical diagnosis. Although many techniques exist for realizing high speed two-photon imaging in the lateral directions, the axial imaging speed is still often limited by the slow mechanical scanning of the objective lens or the specimen, presenting a significant challenge for monitoring fast biological processes at multiple depths as well as in 3D and the development of miniature endoscopy. To overcome this limitation, here a new spectrally encoded two-photon imaging technique is proposed using band-shifting imaging probes, which can enable parallel axial imaging. Specifically, different excitation wavelengths are focused onto different axial positions (through purposely introduced chromatic aberration) to excite two-photon fluorescence from the band-shifting imaging probes, which shift the emission band when the excitation wavelength varies. As such, the fluorescence signals at different axial positions are spectrally encoded to exhibit different spectral bands, and can thus be imaged in parallel by using a spectrometer or arrayed wavelength-resolving detectors. The proposed band-shifting imaging probes will be synthesized, optimized, and used for cellular labeling. Systematic characterization on the molecular structures, molecular weight, photophysical and two-photon properties will be performed. The proposed chromatic two-photon imaging system will be designed, developed, and optimized. System metrics including the mapping relationship between the axial position and fluorescence band shift, the axial imaging range, and spatial resolutions will be characterized. The proposed method will be demonstrated and validated by performing imaging of cells and tissue phantoms as well as by in vivo imaging studies of a colorectal cancer xenograft model.

使用带转换成像探针的色度两光子荧光显微镜 两光子荧光显微镜已成为最重要的成像方式之一 生物学研究和医学诊断。尽管存在许多技术来实现高速两光孔 在横向方向上成像，轴向成像速度仍然通常受到缓慢的机械扫描的限制 目标镜头或样品的构成，对监测快速生物学的挑战提出了重大挑战 多个深度以及3D的过程以及微型内窥镜检查的发展。克服这一点 限制，这里提出了一种新的频谱编码的两光子成像技术 成像探针，可以实现平行的轴向成像。具体而言，不同的激发波长是 专注于不同的轴向位置（通过有目的地引入色差）以激发双光子 从带转移成像探针的荧光，当激发时，它们会移动发射带 波长各不相同。因此，将不同轴向位置的荧光信号频谱编码为 表现出不同的光谱带，因此可以使用光谱仪或阵列并行成像 波长分辨探测器。所提出的带转移成像探针将被合成，优化， 并用于细胞标记。分子结构的系统表征，分子量， 将执行光物理和两光子的特性。提出的色度两光子成像 系统将经过设计，开发和优化。系统指标，包括映射关系 在轴向位置和荧光带移位之间，轴向成像范围和空间分辨率将是 特征。提出的方法将通过进行细胞的成像和 组织幻象以及结直肠癌异种移植模型的体内成像研究。