Leica Stellaris 8 confocal and 2-photon microscope

Project Summary/Abstract We are requesting a Leica Stellaris 8 two-photon (2P) and conventional confocal hybrid microscope system. This instrument is required to maintain and expand biomedical research programs at the University of Montana (UM). It will be integrated into the BioSpectroscopy Core Research Laboratory (BCRL), which is a part of the NIH CoBRE funded Center for Biomolecular Structure and Dynamics (CBSD) at UM. The proposed instrument will be available to users at UM along with external users at regional institutions. This microscope will advance imaging in the life sciences, expanding the capability to detect tissue-specific features related to disease and development. The BCRL currently provides access to a Zeiss 880 laser scanning confocal microscope. However, this instrument has been in service since 2016 and is not currently meeting the needs of BCRL users. One area of deficiency is limited fluorescence lifetime imaging microscopy (FLIM) capabilities of the Zeiss 880. While the Zeiss 880 can perform FLIM, it relies on the integration of PicoQuant parts and software. This results in the acquisition of FLIM images at 50% below Nyquist resolution limit, the need to manually change and/or purchase bandpass filters to adjust the detected emission window, and outdated functional integration between confocal and FLIM modules. The proposed Leica Stellaris 8 offers an "all-in-one" FLIM setup without the need for third party equipment and software. Leica HyD detectors come equipped with variable bandpass filters and dichroics, reducing user maintenance and the need to purchase additional filters. Another critical need of BCRL users is two-photon (2P) microscopy capability, which is not currently available at UM. Our major users plan to image tissue features defined by second harmonic generation, perform 2P FLIM, or image deep within tissues. These needs are only met by a 2P microscope. Imaging of murine lung, murine brain tissue, and Drosophila brains will be enhanced with the acquisition of the proposed microscope. In addition, several technical innovations in conventional confocal microscopy will be advantageous to our users. These include but are not limited to faster acquisition, on-the-fly deconvolution, near-IR detection capabilities, spectral unmixing, and background removal. Thus, a combined 2P and confocal system is sought to meet current user needs, attract new users, and provide state-of-the-art equipment to support NIH funded research. The acquisition of the proposed instrument will advance the research programs of eight NIH-funded major users and two users funded through other sources.