In:
PLOS ONE, Public Library of Science (PLoS), Vol. 15, No. 12 ( 2020-12-15), p. e0243319-
Abstract:
Here, we report a rapid and ultra-sensitive detection technique for fluorescent molecules called scanning single molecular counting (SSMC). The method uses a fluorescence-based digital measurement system to count single molecules in a solution. In this technique, noise is reduced by conforming the signal shape to the intensity distribution of the excitation light via a circular scan of the confocal region. This simple technique allows the fluorescent molecules to freely diffuse into the solution through the confocal region and be counted one by one and does not require statistical analysis. Using this technique, 28 to 62 aM fluorescent dye was detected through measurement for 600 s. Furthermore, we achieved a good signal-to-noise ratio (S/N = 2326) under the condition of 100 pM target nucleic acid by only mixing a hybridization-sensitive fluorescent probe, called Eprobe, into the target oligonucleotide solution. Combination of SSMC and Eprobe provides a simple, rapid, amplification-free, and high-sensitive target nucleic acid detection system. This method is promising for future applications to detect particularly difficult to design primers for amplification as miRNAs and other short oligo nucleotide biomarkers by only hybridization with high sensitivity.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0243319
DOI:
10.1371/journal.pone.0243319.g001
DOI:
10.1371/journal.pone.0243319.g002
DOI:
10.1371/journal.pone.0243319.g003
DOI:
10.1371/journal.pone.0243319.g004
DOI:
10.1371/journal.pone.0243319.g005
DOI:
10.1371/journal.pone.0243319.t001
DOI:
10.1371/journal.pone.0243319.s001
DOI:
10.1371/journal.pone.0243319.s002
DOI:
10.1371/journal.pone.0243319.s003
DOI:
10.1371/journal.pone.0243319.s004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2020
detail.hit.zdb_id:
2267670-3
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