Enabling 3D Fluorescence Imaging Under Anaerobic Environments
Abstract
Enabling 3D Fluorescence Imaging Under Anaerobic EnvironmentsFluorescence microscopy is a crucial analytical tool for biological, mineral, and soft material research, allowing high-resolution and fast time-dependent imaging under standard conditions. We are proposing the acquisition of a Spinning Disk Confocal Microscope (SCDM) at Arizona State University (ASU). The proposed SCDM provides capabilities that are not currently available at ASU and will benefit many DoD and non-DoD projects. The SDCM allows for real-time imaging of processes in the sub-second scale, providing an invaluable tool to image fast kinetic processes. Its minimal fluorescence exposure permits longer-term 3-D imaging in biological samples. Many DoD and non-DoD projects involve oxygen-sensitive biological and non-biological samples; handling these samples for imaging to avoid oxygen exposure is particularly challenging. Thus, the SCDM will be placed inside an anaerobic glovebox to allow imaging of anaerobic samples, including anaerobic microorganisms, oxygen-sensitive materials, and reduced minerals. The SDCM will be the only setup at ASU configured for strict anaerobic microscopy. The compact nature of the SDCM is especially advantageous for this setup. Thus, the SCDM will not only be a cost-effective option for 3-D imaging, it will also significantly enhance the imaging capabilities at ASU. The proposal outlines six DoD projects that will directly benefit from the SDCM. These projects not only demonstrate the impact of the SDCM, but also show its versatility. Within these DoD projects, microbiological samples include anode-respiring bacteria (ARB) capable of producing electrical current, which will be imaged by the SDCM to elucidate fast kinetic transport process within biofilms. Other microbiological samples include microorganisms used for bioremediation of solvents and chlorinated compounds, as well as gastrointestinal samples after a microbiotatransfer therapy for children with autism spectrum disorder. In all of these cases, there is a need to image anaerobic microbial communities in an oxygen-free environment. At the interface between microbiology and material science, one project aims to image fluorescent gold nanoparticles produce through protein synthesis in E. coli. Such imaging will allow establishing a rate and location of the cellular nanoparticle synthesis under different growth conditions. TheSDCM will also allow the imaging of thermoset nanocomposites and investigate their thermomechanical properties. Complementary fluorescent dyes incorporated into functionalized nanoparticles (quantum dots and carbon nanotubes) will form F~rster-resonance energy transfer (FRET) pairs, which enable a temporal investigation of nanoparticle proximity and compositeintegrity under thermomechanical studies. Housed within the Biodesign Institute at ASU, the SDCM will be subjected to strict protocols for usage and maintenance that have previously ensured functionality of many shared equipment. A full-time laboratory coordinator will manage usage and schedule maintenance of the SDCM. We expect this instrument to be functional for over 15 years at ASU, providing a unique approach to fast, anaerobic 3D imaging
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Aug 20, 2019
- Source ID
- N000141912531
Entities
People
- César I. Torres
Organizations
- Arizona State University
- Office of Naval Research
- United States Navy