Highly sensitive and rapid fluorescence detection with a portable FRET analyzer

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Title
Highly sensitive and rapid fluorescence detection with a portable FRET analyzer
Author(s)
Haseong Kim; Gui Hwan Han; Y Fu; J Gam; Seung Goo Lee
Bibliographic Citation
Journal of Visualized Experiments, vol. 2016, no. 116, pp. e54144-e54144
Publication Year
2016
Abstract
Recent improvements in Forster resonance energy transfer (FRET) sensors have enabled their use to detect various small molecules including ions and amino acids. However, the innate weak signal intensity of FRET sensors is a major challenge that prevents their application in various fields and makes the use of expensive, high-end fluorometers necessary. Previously, we built a cost-effective, high-performance FRET analyzer that can specifically measure the ratio of two emission wavelength bands (530 and 480 nm) to achieve high detection sensitivity. More recently, it was discovered that FRET sensors with bacterial periplasmic binding proteins detect ligands with maximum sensitivity in the critical temperature range of 50-55 °C. This report describes a protocol for assessing sugar content in commercially-available beverage samples using our portable FRET analyzer with a temperature-specific FRET sensor. Our results showed that the additional preheating process of the FRET sensor significantly increases the FRET ratio signal, to enable more accurate measurement of sugar content. The custom-made FRET analyzer and sensor were successfully applied to quantify the sugar content in three types of commercial beverages. We anticipate that further size reduction and performance enhancement of the equipment will facilitate the use of hand-held analyzers in environments where high-end equipment is not available.
Keyword
BiochemistryFluorescence resonance energy transferFluorometerFood assessmentForster resonance energy transfer (FRET)Issue 116Point-of-care testingPortable deviceSugar content
ISSN
1940-087X
Publisher
Journal of Visualized Experiments
DOI
http://dx.doi.org/10.3791/54144
Type
Article
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
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