IDBR: Development of High-Speed Two-Photon Excitation Fluorescence Microscopy with Chromatically Extended Depth of Focus

该奖项是为了表彰开发更快的双光子荧光显微镜，特别是在轴向扫描速度方面的改进。速度将通过波分复用（或彩色双光子成像）来提高。虽然在提高横向扫描速度方面取得了很大进展，但高速轴向扫描方法仍然缺失。为了在轴向扫描，物镜或试样本身仍然需要缓慢的机械平移，这限制了3D成像速度。因此，现有仪器在3D和x-z或y-z横截面上监测快速生物过程（例如瞬态钙信号）的能力非常有限。该仪器将对3D显微镜这一生物学中最重要和最广泛使用的工具产生重大影响。它将允许研究人员研究快速动力学（例如，膜运输，细胞内或细胞之间的化学波），并提供各种细胞间和细胞内过程的基本理解，如钙成像。除了科学上的好处，该项目还将涉及本科生和研究生。特别鼓励妇女和少数民族学生参与该项目。调查人员将与宾夕法尼亚州立大学的妇女工程等组织密切合作，为代表性不足的群体和K-12学生开发外展教育项目。

This award is for the development of a faster two-photon fluorescence microscope, with improvements especially in the axial scanning speed. Speed will be increased by wavelength division multiplexing (or chromatic two-photon imaging). Although much progress has been made in improving the lateral scanning speed, a high-speed axial scanning method is still missing. Slow mechanical translation of the objective lens or the specimen itself is still required in order to scan in the axial direction, which limits the 3D imaging speed. As a result, existing instruments have very limited capability to monitor fast biological processes (e.g., transient calcium signals) in 3D and in the x-z or y-z cross-section. The proposed instrument will have a significant impact on 3D microscopy, one of the most important and widely used tools in biology. It will allow researchers to study fast dynamics (e.g., membrane trafficking, chemical waves within or between cells) and provide fundamental understanding of various inter- and intra- cellular processes such as calcium imaging. In addition to its scientific benefits, the project will involve undergraduate researchers as well as graduate students. Woman and minority students will be particularly encouraged to be involved in the program. The investigators will work closely with Penn State organizations such as Women in Engineering to develop outreach education programs for under-represented groups as well as K-12 students.