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Energy transfer chemiluminescence for ratiometric pH imaging.

ABSTRACT: Chemiluminescence imaging offers a low background and high sensitivity approach to imaging analytes in living cells and animals. Intensity-based measurements have been developed, but require careful consideration of kinetics, probe localization, and fluctuations in quantum yield, all of which complicate quantification. Here, we report a ratiometric strategy for quantitative chemiluminescence imaging of pH. The strategy relies on an energy transfer cascade of chemiluminescence emission from a spiroadamantane 1,2-dioxetane to a ratiometric pH indicator via fluorescent dyes in Enhancer solutions. Monitoring the pH-dependent changes in chemiluminescence emission at multiple wavelengths enables ratiometric imaging and quantification of pH independent from variations due to kinetics and probe concentration.

SUBMITTER: An W

PROVIDER: S-EPMC5999021 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Energy transfer chemiluminescence for ratiometric pH imaging.

An Weiwei W Mason Ralph P RP Lippert Alexander R AR

Organic & biomolecular chemistry 20180601 22

Chemiluminescence imaging offers a low background and high sensitivity approach to imaging analytes in living cells and animals. Intensity-based measurements have been developed, but require careful consideration of kinetics, probe localization, and fluctuations in quantum yield, all of which complicate quantification. Here, we report a ratiometric strategy for quantitative chemiluminescence imaging of pH. The strategy relies on an energy transfer cascade of chemiluminescence emission from a spi ...[more]

PMID: 29786719

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Project description:DNAzymes have emerged as an important class of sensors for a wide variety of metal ions, with florescence DNAzyme sensors as the most widely used in different sensing and imaging applications because of their fast response time, high signal intensity, and high sensitivity. However, the requirements of an external excitation light source and its associated power increase the cost and size of the fluorometer, making it difficult to be used for portable detections. To overcome these limitations, we report herein a DNAzyme sensor that relies on chemiluminescence resonance energy transfer (CRET) without the need for external light. The sensor is constructed by combining the functional motifs from both Pb2+-dependent 8-17 DNAzyme conjugated to fluorescein (FAM) and hemin/G-quadruplex that mimics horseradish peroxidase to catalyze the oxidation of luminol by H2O2 to yield chemiluminescence. In the absence of Pb2+, the hybridization between the enzyme and substrate strands bring the FAM and hemin/G-quadruplex in close proximity, resulting in CRET. The presence of Pb2+ ions can drive the cleavage on the substrate strand, resulting in a sharp decrease in the melting temperature of hybridization and thus separation of the FAM from hemin/G-quadruplex. The liberated CRET pair causes a ratiometric increase in the donor's fluorescent signal and a decrease in the acceptor signal. Using this method, Pb2+ ions have been measured rapidly (<15 min) with a low limit of detection at 5 nM. By removing the requirement of exogenous light excitation, we have demonstrated a simple and portable detection using a smartphone, making the DNAzyme-CRET system suitable for field tests of lake water. Since DNAzymes selective for other metal ions or targets, such as bacteria, can be obtained using in vitro selection, the method reported here opens a new avenue for rapid, portable, and ratiometric detection of many targets in environmental monitoring, food safety, and medical diagnostics.

Dataset Information

Energy transfer chemiluminescence for ratiometric pH imaging.

Publications

Energy transfer chemiluminescence for ratiometric pH imaging.

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