Fast Multi-Functional 3D Imaging of Cellular Activities in Deep Tissue

深层组织细胞活动的快速多功能 3D 成像

基本信息

批准号：
10861526
负责人：
Jun Ding
金额：
$ 64.6万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10861526
关键词：
3-Dimensional Address Brain Cell Culture Techniques Cells Complex Cyclic AMP Cyclic AMP-Dependent Protein Kinases Detection Development Device or Instrument Development Dimensions Disease Progression Energy Transfer Face Fluorescence Fostering Functional Imaging Holography Image Imaging Device Imaging Techniques Kidney Label Lateral Light Lighting Location Methods Microscopy Mus Neurophysiology - biologic function Organ Organism Phase Phosphotransferases Process Records Resolution Running Scanning Signal Transduction Slice Speed Structure Subcellular structure System Techniques Testing Three-Dimensional Image Three-Dimensional Imaging Time Tissue imaging Tissues Work Zebrafish absorption cellular imaging fluorescence lifetime imaging imaging modality imaging study improved in vivo in vivo imaging industry partner innovation instrument interest kidney cell lens light curve light intensity light microscopy lightspeed microscopic imaging neural neuroimaging novel open source operation ratiometric sensor spatiotemporal stereoscopic submicron tool two-photon voltage

项目摘要

PROJECT SUMMARY We propose to develop and disseminate an advanced 2p Bessel beam light sheet (2pBLS) microscopy instrument for high-speed sub-micron-resolution volumetric imaging in deep tissue. The proposed instrument will have a significantly higher volume imaging speed over a large field of view, maintain a sub-micron diffraction- limited resolution in deep tissue despite the presence of tissue scatter and aberration, and perform simultaneous lifetime imaging with FRET sensors and high-speed light sheet imaging of dynamics intensity-modulated sensors for dual functional imaging in deep tissue. The system will be tested with two imaging studies in zebrafish and mouse respectively. In zebrafish, we will demonstrate live FLIM imaging of genetically encoded FRET sensors in zebrafish kidney and perform an imaging study of Ca2+ and cAMP activities in zebrafish renal cells. In mice, we will demonstrate imaging of neural and glial activities genetically labeled with activity sensors (Ca2+ and GEVI), FLIM-FRET sensors (PKA) and structural markers ex vivo and in vivo.

项目概要我们建议开发和传播先进的 2p 贝塞尔光束光片 (2pBLS) 显微镜用于深层组织高速亚微米分辨率体积成像的仪器。拟议文书将在大视场范围内具有明显更高的体积成像速度，保持亚微米衍射- 尽管存在组织散射和像差，但深层组织的分辨率有限，并且同时执行使用 FRET 传感器进行寿命成像以及动态强度调制传感器的高速光片成像用于深层组织的双功能成像。该系统将通过斑马鱼和分别鼠标。我们将在斑马鱼中演示基因编码 FRET 传感器的实时 FLIM 成像斑马鱼肾细胞中 Ca2+ 和 cAMP 活性的成像研究。在小鼠中，我们将展示用活动传感器（Ca2+ 和 GEVI）基因标记的神经和神经胶质活动的成像， FLIM-FRET 传感器 (PKA) 和离体和体内结构标记。