Accounts of Chemical Research, volume 49, issue 9, pages 1722-1730
Fluorescent Biosensors Based on Single-Molecule Counting.
Publication type: Journal Article
Publication date: 2016-09-01
Journal:
Accounts of Chemical Research
Quartile SCImago
Q1
Quartile WOS
Q1
Impact factor: 18.3
ISSN: 00014842, 15204898
General Chemistry
General Medicine
Abstract
Biosensors for highly sensitive, selective, and rapid quantification of specific biomolecules make great contributions to biomedical research, especially molecular diagnostics. However, conventional methods for biomolecular assays often suffer from insufficient sensitivity and poor specificity. In some case (e.g., early disease diagnostics), the concentration of target biomolecules is too low to be detected by these routine approaches, and cumbersome procedures are needed to improve the detection sensitivity. Therefore, there is an urgent need for rapid and ultrasensitive analytical tools. In this respect, single-molecule fluorescence approaches may well satisfy the requirement and hold promising potential for the development of ultrasensitive biosensors. Encouragingly, owing to the advances in single-molecule microscopy and spectroscopy over past decades, the detection of single fluorescent molecule comes true, greatly boosting the development of highly sensitive biosensors. By in vitro/in vivo labeling of target biomolecules with proper fluorescent tags, the quantification of certain biomolecule at the single-molecule level is achieved. In comparison with conventional ensemble measurements, single-molecule detection-based analytical methods possess the advantages of ultrahigh sensitivity, good selectivity, rapid analysis time, and low sample consumption. Consequently, single-molecule detection may be potentially employed as an ideal analytical approach to quantify low-abundant biomolecules with rapidity and simplicity. In this Account, we will summarize our efforts for developing a series of ultrasensitive biosensors based on single-molecule counting. Single-molecule counting is a member of single-molecule detection technologies and may be used as a very simple and ultrasensitive method to quantify target molecules by simply counting the individual fluorescent bursts. In the fluorescent sensors, the signals of target biomolecules may be translated to the fluorescence signals by specific in vitro/in vivo fluorescent labeling, and consequently, the fluorescent molecules indicate the presence of target molecules. The resultant fluorescence signals may be simply counted by either microfluidic device-integrated confocal microscopy or total internal reflection fluorescence-based single-molecule imaging. We have developed a series of single-molecule counting-based biosensors which can be classified as separation-free and separation-assisted assays. As a proof-of-concept, we demonstrate the applications of single-molecule counting-based biosensors for sensitive detection of various target biomolecules such as DNAs, miRNAs, proteins, enzymes, and intact cells, which may function as the disease-related biomarkers. Moreover, we give a summary of future directions to expand the usability of single-molecule counting-based biosensors including (1) the development of more user-friendly and automated instruments, (2) the discovery of new fluorescent labels and labeling strategies, and (3) the introduction of new concepts for the design of novel biosensors. Due to their high sensitivity, good selectivity, rapidity, and simplicity, we believe that the single-molecule counting-based fluorescent biosensors will indubitably find wide applications in biological research, clinical diagnostics, and drug discovery.
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Ma F. et al. Fluorescent Biosensors Based on Single-Molecule Counting. // Accounts of Chemical Research. 2016. Vol. 49. No. 9. pp. 1722-1730.
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Ma F., Li Y., Tang B., Zhang C. Fluorescent Biosensors Based on Single-Molecule Counting. // Accounts of Chemical Research. 2016. Vol. 49. No. 9. pp. 1722-1730.
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TY - JOUR
DO - 10.1021/acs.accounts.6b00237
UR - https://doi.org/10.1021/acs.accounts.6b00237
TI - Fluorescent Biosensors Based on Single-Molecule Counting.
T2 - Accounts of Chemical Research
AU - Ma, Fei
AU - Li, Ying
AU - Tang, Bo
AU - Zhang, Chun-yang
PY - 2016
DA - 2016/09/01 00:00:00
PB - American Chemical Society (ACS)
SP - 1722-1730
IS - 9
VL - 49
SN - 0001-4842
SN - 1520-4898
ER -
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@article{2016_Ma,
author = {Fei Ma and Ying Li and Bo Tang and Chun-yang Zhang},
title = {Fluorescent Biosensors Based on Single-Molecule Counting.},
journal = {Accounts of Chemical Research},
year = {2016},
volume = {49},
publisher = {American Chemical Society (ACS)},
month = {sep},
url = {https://doi.org/10.1021/acs.accounts.6b00237},
number = {9},
pages = {1722--1730},
doi = {10.1021/acs.accounts.6b00237}
}
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Ma, Fei, et al. “Fluorescent Biosensors Based on Single-Molecule Counting..” Accounts of Chemical Research, vol. 49, no. 9, Sep. 2016, pp. 1722-1730. https://doi.org/10.1021/acs.accounts.6b00237.