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

描述(申请人提供)：厚组织中的三维活体成像在细胞生物学中越来越重要。人们想要了解细胞在其自然环境中的情况，跟踪一组细胞的行为，并观察移动细胞在细胞基质中的移动。不幸的是，能够成像亚细胞特征的高分辨率光学显微镜被设计成只能成像盖片下的第一微米。随着成像平面向样品中移动得更深，像差会迅速退化图像。这些像差是由物镜浸泡介质和样品安装介质之间的折射率失配(球面像差)和样品本身的折射率变化(样品诱导的像差)引起的。自适应光学(AO)是一种在光学成像中纠正这些像差的很有前途的技术。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，光学像差得到纠正。在光学天文学中，光学光学系统已成功地用于校正由地球大气引起的波前像差。在这项研究项目中，自适应光学技术将被结合到高分辨率广场显微镜中，以实现在盖子下数十微米的高分辨率生物样本的三维成像。将建造单独的显微镜来校正球面像差和样品诱导的像差，并将研究用于生物样品的波前传感器。然后，将设计一种最终的显微镜，既能纠正球面像差，也能纠正样品引起的像差。这项研究可能会对活体组织的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依赖于我们对细胞生物学的理解的科学进步，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.