Enabling high-resolution imaging deep in live tissue with adaptive optics

描述（由申请人提供）：厚组织中的三维实时成像在细胞生物学中越来越重要。人们希望了解自然环境中的细胞，跟踪通过一组细胞的行为，并观察移动的细胞在细胞基质中移动。不幸的是，高分辨率的光学显微镜，能够成像亚细胞的特点，被设计成图像只有第一微米以下的盖玻片。随着成像平面更深地移动到样品中，像差迅速降低图像。这些像差是由物镜浸没介质和样品安装介质之间的折射率失配（球面像差）和样品本身内的折射率变化（样品引起的像差）引起的。自适应光学（AO）是一种在光学成像中显示出很大希望的校正这些像差的技术。AO通过用波前传感器测量波前然后用可变形反射镜校正波前来校正光学像差。在光学天文学中，AO已成功地用于校正由地球大气层引起的波前像差。对于这个研究项目，自适应光学技术将被纳入高分辨率宽视场显微镜，以允许活的生物样品的三维成像在高分辨率下的盖玻片几十微米。将建造单独的显微镜来校正球面像差和样品引起的像差，并将研究用于生物样品的波前传感器。然后，最终的显微镜将被设计，纠正球面和样品引起的像差。这项研究可能会对活体组织荧光成像的分辨率和灵敏度产生巨大影响。医学和公共卫生的进步取决于我们对细胞生物学的理解，包括对活组织中细胞行为的理解。具有自适应光学技术的显微镜将能够深入活体组织进行高分辨率成像，这将有助于回答有关细胞行为的重要问题。

DESCRIPTION (provided by applicant): Three-dimensional live imaging in thick tissue is increasingly important in cell biology. One would like to understand cells in their natural context, follow behaviors traveling through a group of cells and watch mobile cells move through the cellular matrix. Unfortunately, high-resolution optical microscopes, capable of imaging subcellular features, are designed to image only the first micron below the coverslip. As the imaging plane is moved deeper into the sample, aberrations rapidly degrade the image. These aberrations are caused by the refractive index mismatch between the objective immersion medium and the sample mounting medium (Spherical aberrations) and refractive index variations within the sample itself (Sample-induced aberrations). Adaptive Optics (AO) is a technology that shows great promise for correcting these aberrations in optical imaging. AO corrects optical aberrations by measuring the wavefront with a wavefront sensor and then correcting the wavefront with a deformable mirror. AO has been used with great success in optical astronomy for correcting the wavefront aberrations caused by the earth's atmosphere. For this research project, adaptive optics technology will be incorporated into high-resolution wide-field microscopes to allow three-dimensional imaging of living biological samples at high-resolution many tens of microns below the coverslip. Separate microscopes will be built to correct spherical aberrations and sample- induced aberrations, and research will be done into wavefront sensors for biological samples. Then a final microscope will be designed that corrects both spherical and sample-induced aberrations. This research could have a tremendous impact on the resolution and sensitivity of fluorescence imaging into live tissue. Advances in medical science and public health depend upon scientific advances in our understanding of cell biology, including understanding the behavior of cells in living tissue. A microscope with adaptive-optics technology will enable high-resolution imaging deep into live tissue which will help answer important questions about cell behavior.