By Whitney Harder
(April 25, 2016) — A new instrument to be developed by ÌÇÐÄvlog¹Ù·½Èë¿Ú researchers will overcome current limitations in fluorescence microscopy and could accelerate basic scientific discoveries. The multimodal and cost-effective imaging and data collection platform is being funded by a three-year, $589,250 National Science Foundation (NSF) grant from the Instrument Development for Biological Research (IDBR) program.
Total internal reflection fluorescence (TIRF) microscopy and fluorescence correlation spectroscopy (FCS) have become essential tools to understand biochemical and cellular processes. But these tools are limited — they are often highly labor intensive and thus have been primarily restricted to single sample analysis followed by costly manual data processing.
UK Department of ÌÇÐÄvlog¹Ù·½Èë¿Ú Assistant Professors Jason DeRouchey and Chris Richards, as well as Professor , will develop high throughput versions of both techniques under a single platform, giving researchers the ability to simultaneously scan multiple samples.
"Our instrument will be the first-of-its-kind; currently, there are no instruments capable of performing high throughput TIRF and only a few custom instruments capable of high throughput FCS," DeRouchey said.
DeRouchey noted the new instrument could also have direct applications in phenotypic screening and lead to entirely new methods for high throughput screening in vivo and in vitro.
DeRouchey and Richards will use their expertise in single molecule spectroscopy to develop the hardware for the instrument while Yang will develop an open source data analysis platform, Open Fluorescence Spectroscopy (OpenFS), made freely available online. OpenFS will provide an alternative to costly and proprietary imaging software often tied to a particular device.
"Currently in single molecule fluorescence, one of the greatest barriers to more users applying these methods is not the instrumentation (many people have access to confocal microscopes), but rather understanding how to treat the data collected," DeRouchey said.
While some analysis packages are currently available, the UK team aims to be the first to create a modular, cost-free software package that can integrate both instrument control components and data analysis software. Working with the , the team plans to host the software package in an interactive forum where users can pick and choose modules developed by the UK team for a range of applications, as well as upload their own modules to share with the community.
"Our hope is that OpenFS can therefore compete or replace the fragmented and often proprietary analysis software currently available," DeRouchey said.
Research enabled by this project will span the disciplines of chemistry, physics, molecular biology and engineering to answer fundamental questions in biology. The multidisciplinary nature of the project will also create unique training and educational opportunities for undergraduate, graduate and post-doctoral researchers at UK.