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Related Experiment Video

Updated: Nov 25, 2025

Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions

Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions

Published on: June 14, 2016

Scanning single-molecule counting system for Eprobe with highly simple and effective approach.

Takeshi Hanami^1,2, Tetsuya Tanabe^1,2,3, Takuya Hanashi^1,2,3

¹Genetic Diagnosis Technology Unit, RIKEN Center for Integrative Medical Science, Yokohama, Kanagawa, Japan.

|December 15, 2020

Summary

We developed scanning single molecular counting (SSMC), a rapid and ultra-sensitive method to detect fluorescent molecules by counting them individually. This technique achieves high sensitivity for detecting target nucleic acids, even for challenging biomarkers like miRNAs.

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Last Updated: Nov 25, 2025

Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions

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Integrated Cell Manipulation Platform Coupled with the Single-probe for Mass Spectrometry Analysis of Drugs and Metabolites in Single Suspension Cells

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Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

Area of Science:

Analytical Chemistry
Biotechnology
Molecular Diagnostics

Background:

Accurate detection of low-concentration molecules is crucial for diagnostics.
Existing methods often require amplification or complex statistical analysis.
Developing sensitive and rapid detection techniques remains a challenge.

Purpose of the Study:

To introduce a novel, rapid, and ultra-sensitive detection technique for fluorescent molecules.
To demonstrate the capability of counting single molecules in solution without amplification.
To establish a highly sensitive nucleic acid detection system.

Main Methods:

Scanning Single Molecular Counting (SSMC) utilizes a fluorescence-based digital system.
Noise reduction is achieved by conforming signal shape to excitation light intensity via circular scanning.
Single fluorescent molecules diffuse freely through the confocal region and are counted individually.

Main Results:

Detection of 28 to 62 aM fluorescent dye within 600 s.
Achieved a high signal-to-noise ratio (S/N = 2326) for 100 pM target nucleic acid.
Demonstrated amplification-free detection using hybridization-sensitive fluorescent probes (Eprobe).

Conclusions:

SSMC combined with Eprobe offers a simple, rapid, and highly sensitive nucleic acid detection system.
The method is promising for detecting difficult-to-amplify biomarkers such as miRNAs and short oligonucleotides.
This approach eliminates the need for amplification and complex statistical analysis.