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Title 

Highly sensitive and rapid fluorescence detection with a portable FRET analyzer

Authors 

Haseong KimGui Hwan HanY FuJ GamSeung Goo Lee

Publisher 

Journal of Visualized Experiments (JoVE)

Issue Date 

2016

Citation 

Journal of Visualized Experiments, vol. 2016, no. 116, pp. e54144-e54144

Keywords 

BiochemistryFluorescence resonance energy transferFluorometerFood assessmentF?rster resonance energy transfer (FRET)Issue 116Point-of-care testingPortable deviceSugar content

Abstract 

Recent improvements in F?rster 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.

ISSN 

1940-087X

Link 

http://dx.doi.org/10.3791/54144

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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