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

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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.