Abstract Details

Title Continuous Laser Induced Fluorescence Spectroscopy (CLIFS) Technique for Screening Drugs by Assessing the Metabolic Effects in Real-Time

Topic Cerebrovascular Disease and Interventional Neurology

Presentation(s) P04 - (-)

Poster/Presentation
Number 065

Objective

Background BACKGROUND: Currently, there is no method to measure in real-time the effect of various therapies on the cellular metabolism. We propose to use CLIFS as a potential method to assess the same. CLIFS allows the continuous monitoring of multiple fluorescent metabolites (400 - 650 nm) in-vitro / in-vivo.

Design/Methods DESIGN/METHODS: In this study, we focus on using the NADH fluorescence by exciting the ex-vivo rabbit brain tissue with pulsed ultraviolet laser (350 nm) and recording the fluorescence emission at two wavelength bands between 400-450 nm & 451-480 nm, allowing for differential measurements of bound and free form of NADH, respectively. While the fluorescence is being recorded from the brain tissue sample, a drug in various concentrations is added to the medium. Changes in the NADH fluorescence level are recorded and analyzed.

Results RESULTS: We found that rotenone (50 nanoM - 50 microM), a known inhibitor of electron transport chain increased the overall NADH fluorescence indicating accumulation of the metabolite in the cells. Additionally, it was observed that the bound form of NADH increased to an high extent that the free form. We also noted that rotenone has a rapid effect on the cellular metabolism. We will also present our preliminary results in using CLIFS for screening a library of potential neuro-protective drugs, based on the hypothesis that NADH levels correlates to the energy metabolism within the cell.

Conclusions CONCLUSIONS: CLIFS allows us to measure effects of neuro-protective drugs in real-time (300 ms interval) and has demonstrated a potential as a new screening method to assess metabolic effects of drugs.

Authors/Disclosures
Pramod Butte PRESENTER	No disclosure on file
Paul A. Lapchak, PhD, FAHA (Cedars-Sinai Medical CenterAdvanced Health Sciences Pavilion)	No disclosure on file
	No disclosure on file

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