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.

使用带移成像探针的彩色双光子荧光显微术 双光子荧光显微镜已成为最重要的成像模式之一， 生物学研究和医学诊断。尽管存在许多用于实现高速双光子的技术， 在横向方向上成像时，轴向成像速度仍然常常受到缓慢的机械扫描的限制 物镜透镜或样本的，提出了一个重大的挑战，监测快速生物 在多个深度以及在3D和微型内窥镜的发展过程。为了克服这个 限制，这里提出了一种新的光谱编码双光子成像技术，使用带移 成像探头，其能够实现平行轴向成像。具体地，不同的激发波长是 聚焦到不同的轴向位置（通过故意引入的色差）以激发双光子 来自带移成像探针的荧光，当激发时， 波长变化。因此，在不同轴向位置处的荧光信号被光谱编码， 显示出不同的光谱带，因此可以通过使用光谱仪或阵列 波长分辨探测器所提出的带移成像探针将被合成、优化， 并用于细胞标记。对分子结构、分子量、 将进行光物理和双光子性质的研究。彩色双光子成像 系统将被设计，开发和优化。包括映射关系的系统度量 在轴向位置和荧光带偏移之间，轴向成像范围和空间分辨率将是 表征了所提出的方法将通过对细胞进行成像来证明和验证， 组织模型以及结肠直肠癌异种移植模型的体内成像研究。