Enabling Enhanced Biofilm Imaging for Department of Defense-Related Research at Montana State University and the Northwestern United States
Abstract
Established at Montana State University (MSU) in 1990, the Center for Biofilm Engineering (CBE) is the worldÕs first, largest, and best-known biofilm research center. Biofilms are aggregates of microorganisms that are embedded within a matrix of self-assembled extracellular polymeric substances and adhere to each other and/or surfaces. Microbial biofilms are present in almost all known habitats, including nature, industry, and the human body, and often exist as multi-species communities with extreme complexity. The investigation of biofilms is well justified, and few other research areas have far-reaching implications for water, food, energy, national security, and health. All currently DoD-funded projects at the CBE rely on optical microscopy (OM) (1) 3D Printing of Microbial Communities for Optimal Resource Processing, (2) Development of Robust Microbial Communities through Engineered Biofilms, and (3) VIPER: Viral Interdiction through Population Engineering and Restructuring. Real-time imaging capabilities are essential for these projects and OM is well suited for biofilm studies and the required applications. However, technical limitations of our current ~25-year old widefield and 10-year old confocal scanning laser microscope (CSLM) (slow scan/acquisition speed, and consequently photodamage from longer dwell times) hinder researchers from conducting high resolution and high throughput live cell imaging. The proposed instrument is optimally configured for cutting-edge, real-time, high-sensitivity imaging of complex and intact biological samples and thus, ideally suited for biofilm systems. With the proposed instrument we can build and sustain competitive research capabilities for CBE, MSU, and our collaborators to advance DoD-relevant research. We propose to acquire a uniquely configured THUNDER Live Cell epifluorescence widefield microscope from Leica Microsystems, Inc. In the past, imaging biofilms has relied heavily on CSLMs due to the ability to provide 3D information of hydrated biofilms noninvasively. Although epifluorescence widefield OM would be better suited, high background signals have limited applications. The proposed THUNDER imager uses computational clearing to remove background signal in real time. Acquisition speeds of the proposed system are nearly five times faster than a CSLM, and experiments that would take hours to visualize on a CSLM can be accomplished in minutes. The proposed widefield microscope would provide 1.) improved Ôunder-microscopeÕ incubation capabilities that will improve live-cell imaging and relevance, 2.) reduced phototoxicity and photobleaching, which is essential to maintain cell viability during live imaging, 3.) increased image acquisition speed, essential for high throughput imaging and interfacing OM with modern microfluidic and cell biology approaches, and 4.) increased imaging depth via real time computational removal of out of focus background. The acquisition of the THUNDER Live Cell epifluorescence widefield microscope will provide a critical addition to our imaging facility and MSUÕs research infrastructure to enhance research on currently funded DoD projects and as well as related projects. The requested equipment will interface with current CBE microfluidic platforms and other correlative OM capabilities. The proposed optical microscope will also enable the expansion into DoD areas of interest including viral and bacterial infections, kidney stone development, and metal corrosion. The CBE is a student-centered, interdisciplinary center that bridges fundamental and applied research, and the microscope facility is a centerpiece of the student research experience. The requested addition to our imaging facility will increase our ability to remain competitive in research areas of federal interest as well as broaden our ability to provide access to, and training for, technologically advanced research instruments.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jun 25, 2021
- Source ID
- W911NF2110066
Entities
People
- Matthew W. Fields
Organizations
- Army Contracting Command
- Montana State University
- United States Army