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

描述（由申请人提供）：厚组织的三维实时成像在细胞生物学中越来越重要。人们想要了解细胞在自然环境中的情况，跟踪一组细胞的行为，观察移动细胞在细胞基质中的移动。不幸的是，高分辨率光学显微镜，能够成像亚细胞特征，被设计成只能成像盖盖下面的第一微米。随着成像平面深入样品，像差会迅速降低图像的质量。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不匹配（球面像差）和样品本身的折射率变化（样品诱导像差）引起的。自适应光学技术（AO）是一种很有希望用于光学成像中校正这些像差的技术。AO通过使用波前传感器测量波前，然后使用可变形镜校正波前来校正光学像差。AO在光学天文学中用于校正地球大气引起的波前像差取得了巨大的成功。在这个研究项目中，自适应光学技术将被整合到高分辨率宽视场显微镜中，以允许在盖盖下几十微米的高分辨率下对活体生物样品进行三维成像。将建立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。然后设计一个最终的显微镜来校正球面像差和样品引起的像差。这项研究将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的进步，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对活组织进行深入的高分辨率成像，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.