Progress 09/01/24 to 08/31/25

Outputs
Target Audience:The Cellular Analysis Core (CAC) facility that manages the ZE5 Cell Analyzer is a research core facility open to all researchers at Montana State University. Over the past year, the CAC moved from the Department of Microbiology and Cell Biology to MSU's Vice President for Research and Economic Development's Core Facilities Program, which now providing financial and administrative support for our facility. This move has increased the visibility of the core facility, leading to an increase in users of all core instruments including the ZE5 Cell Analyzer. The target audience for this project are undergraduate and graduate students, postdoctoral fellows, faculty members and staff scientists from several of MSU's life sciences departments. In addition, our core facility is also open to external users. Researchers from Montana institutions that are part of the NIH-funded MT INBRE network can use the Cellular Analysis Core including the ZE5 for reduced user fees. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided training in flow cytometry techniques and technology to undergraduate students, graduate students, postdoctoral fellows and other researchers. A total of 16 investigators were able to gain research experience with the new ZE5 flow cytometer. Over the past year, nine new students received hands-on training on the ZE5 Cell Analyzer by our core facility staff. In addition, our Cellular Analysis Core hosted a seminar about advanced flow cytometry techniques for core users and other interested investigators in February 2025. We also will advertise our core facility and flow cytometry equipment including the ZE5 Cell Analyzer by giving a seminar talk at MSU's first Core Day, which will take place on October 3rd, 2025. How have the results been disseminated to communities of interest?Information about the ZE5 Cell Analyzer is provided through our Cellular Analysis Core Website, which is now linked to MSU's Core Facilities Program website. Our website was recently updated to include up-to-date infromation for our users on instrument capacity,access and facility fees and to conform to a standard MSU core facility format. In addition, data collected on our instrument have been shared with the public through at least two published manuscripts. We also advertised our core facility to users during the annual new faculty orientation event. As part of the internal process to become a VPR approved core facility, Dr. Bimczok presented information on the core facility and all our key instruments to the MSU research council, which is composed of faculty members across multiple colleges and academic departments, in November 2024. What do you plan to do during the next reporting period to accomplish the goals?In the next funding period, we will continue to provide routine instrument maintenance and user support for the ZE5 Cell Analyzer, so that we can keep the instrument in good working condition while also supporting increased research output. To increase instrument use, we will provide regular email updates to facility users, and we will participate in MSU's first Core Day, which will take place on October 3rd. The Core Day will involve a seminar about our core facility and available services and instruments and "open house" facility tours. The MSU Core Day is being widely advertised to MSU students, faculty, and other investigators, as well as to local biotechnology companies. We also will include a module on flow cytometry methods in the graduate course in Advanced Immunology, which is offered every other year including in Spring 2026. This course will enable graduate students to gain an in depth understanding about flow cytometry technology and applications, and about the instruments including the ZE5 that are currently available at MSU. MSU's Core Facility program has recently obtained a contract for a new core facility management software, Stratocore, will is being rolled out in Fall 2025. This software will facilitate and streamline user management, instrument signup, and invoicing. .

Impacts
What was accomplished under these goals? With this instrument award, we purchased a 30-parameter, high throughput BioRad ZE5 Cell Analyzer, as described in the proposal. The ZE5 Cell Analyzer is a state-of-the art flow cytometer that features five lasers, 27 fluorescence channels, a small particle analyzer, a universal plate and tube loader, and a robust and user-friendly design. The ZE5 Cell Analyzer was installed in the MSU Cellular Analysis Core in November 2023 and has since been open for users. Since the submission of the last progress report, the ZE5 has been used by 16 researchers from eight laboratories from three MSU departments and one local biotechnology company. ZE5 usage has increased significantly during the last reporting period, with a total of 342 hours reported between September 2024 and July 2025, compared to around 100 hours between November 2023 and August 2024. To date, at least two publications from MSU researchers include data collected on the ZE5. During the past year, our Cellular Analysis Core, where the ZE5 Cell Analyzer represents our newest and most versatile flow cytometer, successfully completed the competitive process to become an official "RED" core under the umbrella of MSU's Vice President for Research and Economic Development's Core Facilities Program. Our core facility is now receiving significant administrative and financial support from the University. Participation in MSU's RED core program has already led to increased user numbers and multiple opportunities to promote our facility, such as a new website linked to the Core Facility Program, participation in the first MSU "Core Day", and promotion of the core at the new faculty orientation. A new centralized software solution for instrument scheduling and invoicing will be rolled out over the next few months. The Cellular Analysis Core also is a key component of the Research Core for an NIH Center of Biomedical Research Excellence (COBRE) proposal that was re-submitted in January 2025 and that has received a potentially fundable score. If selected for funding, the COBRE will provide support for core facility personnel and for instrument maintenance contracts for up to 15 years.

Publications

• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Innovative Methodology for Antimicrobial Susceptibility Determination in Mycoplasma Biofilms. Jacobson BT, DeWit-Dibbert J, Selong ET, Quirk M, Throolin M, Corona C, Sonar S, Zanca L, Schwarz ER, Bimczok D. Microorganisms. 2024 Dec 20;12(12):2650. doi: 10.3390/microorganisms12122650. PMID: 39770853
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2025 Citation: TLR2 or TLR4 Stimulation Induces Transmembrane TNF-Driven Priming in Macrophages Which Results in Improved Clearance of a Subsequent Staphylococcus aureus Infection. Luu AM, Hatton AA, Gattiker J, Shepardson KM, Hall MN, Benson E, Bimczok D, Rynda-Apple A. J Innate Immun. 2025;17(1):302-319. doi: 10.1159/000546011. Epub 2025 May 28. PMID: 40435995

Progress 09/01/23 to 08/31/24

Outputs
Target Audience:The Cellular Analysis Core facility that manages the ZE5 Cell Analyzer is a research core facility open to all researchers at Montana State University. The target audience for this project are undergraduate and graduate students, postdoctoral fellows, faculty members and staff scientists in the Department of Microbiology and Cell Biology and other life sciences departments. In addition, our core facility is also open to external users. Researchers from Montana institutions that are part of the NIH-funded MT INBRE network can use the Cellular Analysis Core including the ZE5 for reduced user fees. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided training in flow cytometry techniques and technology to undergraduate students, graduate students, postdoctoral fellows and other researchers through hands-on and seminar training sessions offered by the instrument manufacturer BioRad and a methods seminar talk by Dr. Bimczok at the Microbiology and Cell Biology seminar series. In addition, MSU research trainees are now using the ZE5 Cell Analyzer for their various research projects and are thus gaining valuable flow cytometry experience with a state-of the art instrument. In particular, MSU trainees have been using the small particle analyzer and developing multi-parameter panels, two applications that were not possible previously. How have the results been disseminated to communities of interest?Following the installation of the BioRad ZE5 Cell Analyzer in November 2023, a training seminar was held that provided information on the instrument. In addition, hands-on user training sessions were also provided by a BioRad representative, which were attended by >15 MSU researchers from six labs. Leading up to the instrument installation and training sessions, PI Dr. Bimczok gave a presentation at the Microbiology and Cell Biology Seminar Series in October 2023, where she provided information on the Cellular Analysis Core Facility, relevant methods, technologies, and applications for flow cytometry, and the specific features and of the new ZE5 Cell Analyzer. This seminar was attended by >30 individuals and enabled us to spread the word about flow cytometry capacity at MSU and particularly the high-throughput, high-parameter ZE5. The ZE5 Cell Analyzer is now listed on the MSU Cellular Analysis Core Website and is available to trained users at all career stages, with new user training provided by Core Manager Mr. A. Sebrell. What do you plan to do during the next reporting period to accomplish the goals?In the next funding period, we will continue to provide routine instrument maintenance and user support for the ZE5 Cell Analyzer, so that we can keep the instrument in good working condition while also supporting increased research output. To increase instrument use, we will further update our website, provide email updates to facility users, and provide information about flow cytometry in internal seminars. We also will include a module on flow cytometry methods in the graduate course in Advanced Immunology, so that graduate students will gain an in depth understanding about flow cytometry technology and applications, and about the instruments including the ZE5 that are currently available at MSU. During the next funding period, we also anticipate that we will approval for our core facility as an official "RED" core facility, which will provide increased financial and administrative support for the core facility from the Vice President for Research and Economic Development Office (VPRED). Our internal application for VPRED approval is currently under consideration. A designation of our core facility as a "RED" core facility will also greatly enhance the visibility of the core, leading to an increased number of users. This will ultimately benefit the current project, as increased facility funding through university funds and user fees will enable us to maintain the instrument, perform repairs if needed, and possibly invest in instrument upgrades such as a robotic plate loader in the future.

Impacts
What was accomplished under these goals? With this instrument award, we purchased a30-parameter, high throughput BioRad ZE5 Cell Analyzer, as described in the proposal. The ZE5 Cell Analyzer is a state-of-the art flow cytometer that features five lasers, 27 fluorescence channels, a small particle analyzer, a universal plate and tube loader, and a robust and user-friendly design. The ZE5 Cell Analyzer was installed in the MSU Cellular Analysis in November 2023, with hands-on and seminar-style training sessions held during the same month. The instrument has been available to internal and external users since then and has been used for 100> hours, with an increasing trend. To make the instrument accessible for MSU researchers, we have updated our core website and have linked the new ZE5 Cell Analyzer to our Cellular Analysis Facility in FluoroFinder, a free online resource that enables scientists to design and optimize antibody panels for specific flow cytometers. In addition to the ZE5 Analyzer, this award also enabled us to purchase a Millipore Ultrapure Water Purification system. This system is now providing clean water to run the ZE5 Cell Analyzer instrument, providing cost savings and contributing to the sustainability of the instrument and our core facility. Overall, the new ZE5 Cell Analyzer has significantly enhanced our flow cytometry capacity at MSU. With this new instrument in place, we have submitted an internal application to obtain approval and support for the Cellular Analysis Core by MSU's Vice President for Research (VPR). This application is currently undergoing review. If approved, our facility will receive financial and administrative support and will be eligible for internal funding opportunities to maintain, upgrade and or repair core instruments including the ZE5. In addition, designation of our core facility as a VPR core will significantly enhance the profile and visibility of our facility at our university. In addition, The Cellular Analysis Core also is a key component of the Research Core for an NIH Center of Biomedical Research Excellence (COBRE) proposal that was submitted in January 2024 and that is currently being considered for funding. If selected for funding, the COBRE will provide support for core facility personnel and for instrument maintenance contracts for up to 15 years.

Publications

• Type: Journal Articles Status: Under Review Year Published: 2024 Citation: B. Tegner Jacobson, Jessica DeWit, Eli T. Selong, McKenna Quirk, Michael Throolin, Chris Corona, Sobha Sonar, LaShae Zanca, Erika R. Schwarz, Diane Bimczok Innovative methodology for antimicrobial susceptibility de-termination in M. bovis biofilms Microorganisms. In revision.
• Type: Other Status: Other Year Published: 2023 Citation: D. Bimczok. Leveraging the cellular analysis core for high throughput, multi-dimensional cell characterization. MSU Microbiology and Cell Biology Seminar Series, October 2023, Bozeman, MT