PCR-Based Analysis of a Common Fluorescein Derivative Using Broadband Spectroscopy Platforms

PCR-based testing has rapidly increased in popularity and development since the start of the COVID-19 pandemic, and many new techniques are leveraging the latest performance advancements offered by the spectroscopy systems used to analyze PCR tubes. Guenther evaluates the common fluorescent reporter dye 5(6)-carboxy-2,'7'-dichlorofluorescein (CDCF) in a PCR tube setting using broadband spectroscopy platforms. CDCF is a marker used in the study of cellular antioxidant activity (CAA) which has seen increased focus on the cancer-preventing abilities of antioxidants to inhibit intracellular free radical damage.

This work demonstrates how fluorescence efficiencies can be optimized by addressing optical bench parameters, light source options, and fiber configurations. Furthermore, these optimized PCR-tube readings assess detection limits as a function of marker concentration and extent of quenching. Deeper limits of detection allow greater visibility into the mechanisms that drive these intracellular processes.

About the presenter
Derek Guenther has a bachelor’s degree in chemical engineering, with a minor in inorganic chemistry, from the University of South Florida (USF). His work at the USF Institute for Environmental Studies focused on novel silica-based algaecides targeting invasive species. Guenther’s strengths and passions lie in new chemical sensor development and spectral numerical processing. He has spent over 14 years at Ocean Insight developing and supporting the optical oxygen, pH, and SERS-based sensor products. Today he works with the lab services team helping customers achieve their goals.