Recipient Organization
UNIVERSITY OF SOUTH ALABAMA
307 UNIVERSITY BLVD
MOBILE,AL 36688
Performing Department
(N/A)
Non Technical Summary
Total Internal Reflection Fluorescence (TIRF) microscopyis a special type of imaging that lets us see what is happening right at the surface of a cell. Most microscopes can only capture activity deeper inside the cell, but TIRF works like a spotlight that only shines on the very thin layer at the cell's outer edge. This makes it possible to watch molecules move, interact, and change in real time with incredible detail.In this proposal, we are requesting a high-resolution CSU-W1 E-TIRF system, which offers state-of-the-art imaging speed and resolution, allowing us to cellular dynamics at high spatiotemporal resolution. This new microscope will replace the outdated system and be housed in the Biology Department, where it will be accessible to faculty, students, and collaborators across multiple departments and colleges. By doing so, it will greatly expand training, teaching, and research opportunities, and support groundbreaking discoveries in areas such as plant biology and infectious disease.
Animal Health Component
20%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
0%
Goals / Objectives
At the University of South Alabama, there is no existing TIRF-confocal system that fits our research needs. The USA Department of Biology currently has one existing Leica confocal microscope which was a transferred asset from the USA Whiddon College of Medicine (COM). However, this system is 25 years old, outdated, and was recently decommissioned. All of our imaging experiments are currently performed at the Bioimaging core at COM.Although there are confocal microscopes available, none of these systems has the capability to perform Total Internal Reflection Fluorescence (TIRF) microscopy imaging to visualize molecular dynamics at the cell membrane. The COM core imaging facility is also heavily used, which means setting up imaging experiments that continuously acquire data for several days or weeks is not permissible. Compared to the current confocal systems at COM, the requested CSU-W1 E-TIRF system is equipped with Ti2-LAPP modular illuminators, high-speed piezo Z-drive, and ORCA Fusion sCMOS camera, which has very high axial resolution (200 nm) and speed (89 frames/sec). The system is uniquely suited for cellular and subcellular localization of fluorophores and quantification of host/pathogen's protein abundance and dynamics at the cell plasma membrane (Appendix 2). The CSU Spinning Disk, combined with Ti2-LAPP modular TIRF allow for unprecedented fast and simultaneous multi-color excitation and detection, even at different penetration depth. We will house this CSU-W1 E-TIRF system in the Imaging Facility at the USA Department of Biology as a replacement for the decommissioned Leica system. This will provide high access and visibility to encourage collaborations, instructional, training and research opportunities for all faculty and student researchers at our four Departments, two Colleges (Arts & Sciences and Medicine), the University of South Alabama, and beyond. The CSU-W1 E-TIRF system is a well-integrated equipment for enabling discoveries in the most critical frontiers of plant biology.
Project Methods
Not relevant (this is an equipment grant)