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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Updated: Dec 30, 2025

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
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High-Fidelity Single Molecule Quantification in a Flow Cytometer Using Multiparametric Optical Analysis.

Lucas D Smith1,2, Yang Liu1,2, Mohammad U Zahid1,2

  • 1Department of Bioengineering , University of Illinois at Urbana-Champaign , Urbana , Illinois 61801 , United States.

ACS Nano
|January 24, 2020
PubMed
Summary
This summary is machine-generated.

Commercial flow cytometers can now detect single nucleic acid molecules, like microRNAs, using rolling circle amplification and multicolor labeling. This breakthrough enables high-throughput, sensitive molecular detection with improved signal-to-noise ratios.

Keywords:
RCAamplificationbiomarkerdiagnosticsmiRNAmicroRNAmultiplexing

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Microfluidic techniques excel at analyzing micron-scale objects but struggle with molecular targets.
  • Current single-molecule methods often involve low-throughput imaging of immobilized molecules.

Purpose of the Study:

  • To adapt commercial flow cytometers for single-molecule nucleic acid detection.
  • To develop a method for sensitive and high-throughput quantification of microRNAs.

Main Methods:

  • Enzymatic extension and dense labeling of microRNAs using rolling circle amplification (RCA).
  • Utilizing multicolor fluorophores to create repetitive nucleic acid products with tunable spectral profiles.
  • Cross-correlating multiparametric optical features for signal amplification.

Main Results:

  • Achieved a 1600-fold amplification in signal-to-background ratio, enabling single-molecule detection.
  • Demonstrated a limit of detection of 47 fM for microRNAs, surpassing PCR-based methods by 100-fold.
  • Enabled discrimination of different microRNA sequences in solution using unique optical barcodes.

Conclusions:

  • Commercial flow cytometry can be repurposed for high-throughput, multiparametric single-molecule analysis.
  • The developed method offers a sensitive and accessible approach for molecular detection.
  • This technology has the potential to be adapted for various molecular amplification techniques.