Abstract

The Time-resolved fluorescence spectroscopy (TR-FS) has the potential to differentiate tumor and normal tissue in real time during surgical excision. In this manuscript, we describe the design of a novel TR-FS device, along with preliminary data on detection accuracy for fluorophores in a mixture. The instrument is capable of near real-time fluorescence lifetime acquisition in multiple spectral bands and analysis. It is also able to recover fluorescence lifetime with sub-20ps accuracy as validated with individual organic fluorescence dyes and dye mixtures yielding lifetime values for standard fluorescence dyes that closely match with published data. We also show that TR-FS is able to quantify the relative concentration of fluorescence dyes in a mixture by the unmixing of lifetime decays. We show that the TR-FS prototype is able to identify in near-real time the concentrations of dyes in a complex mixture based on previously trained data. As a result, we demonstrate that in complex mixtures of fluorophores, the relative concentration information is encoded in the fluorescence lifetime across multiple spectral bands. We show for the first time the temporal and spectral measurements of a mixture of fluorochromes and the ability to differentiate relative concentrations of each fluorochrome mixture in real time.

Highlights

  • The Time-resolved fluorescence spectroscopy (TR-Fluorescence Spectroscopy (FS)) has the potential to differentiate tumor and normal tissue in real time during surgical excision

  • We have previously shown the potential of Time-resolved fluorescence spectroscopy (TR-FS) in distinguishing brain tumor from normal brain tissue in a human clinical trial33

  • We demonstrate that the TR-FS prototype is able to quantify lifetime fluorescence decay with high accuracy by determining the fluorescence lifetime of fluorescence standards as well as organic fluorophores

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Summary

Introduction

The Time-resolved fluorescence spectroscopy (TR-FS) has the potential to differentiate tumor and normal tissue in real time during surgical excision. We used the fluorescence standards Rose Bengal and Rhodamine B, as well as NADH, FAD, Protoporphyrin IX and their mixtures with different relative concentrations.

Results
Conclusion
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