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

基本信息

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
项目类别：
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

项目摘要

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镜双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。尽管存在许多实现高速双光子的技术在横向成像时，轴向成像速度仍然常常受到缓慢的机械扫描的限制物镜或样本的变化，对监测快速生物提出了重大挑战多个深度以及 3D 过程以及微型内窥镜的发展。为了克服这个限制，这里提出了一种使用带移的新光谱编码双光子成像技术成像探头，可以实现平行轴向成像。具体来说，不同的激发波长是聚焦到不同的轴向位置（通过故意引入色差）以激发双光子来自带移成像探针的荧光，当激发时，带移成像探针会移动发射带波长不同。因此，不同轴向位置的荧光信号被光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。所提出的带移成像探针将被合成、优化、并用于细胞标记。分子结构、分子量、将执行光物理和双光子特性。所提出的彩色双光子成像系统将被设计、开发和优化。系统指标包括映射关系轴向位置和荧光带位移之间，轴向成像范围和空间分辨率将是特点。所提出的方法将通过进行细胞成像和组织模型以及结直肠癌异种移植模型的体内成像研究。