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Related Experiment Video

Updated: Aug 6, 2025

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Single-cell HER2 quantification via instant signal amplification in microdroplets.

Xiaoxian Liu1, Yifan Zhu1, Caoxin Li1

  • 1College of Engineering and Applied Sciences, Nanjing University, Jiangsu, 210093, China; Key Laboratory of Intelligent Optical Sensing and Integration of the Ministry of Education, Nanjing University, Jiangsu, 210009, China.

Analytica Chimica Acta
|March 16, 2023
PubMed
Summary

A new droplet microfluidic method offers ultrasensitive, quantitative analysis of human epidermal growth factor receptor 2 (HER2) protein on single cells. This approach improves accuracy and reduces variability for better breast cancer diagnosis and treatment.

Keywords:
Droplet microfluidicsHER2 protein expressionInstant cation exchange signal amplificationQuantitative detectionSingle-cell

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Cancer Research

Background:

  • Accurate quantification of human epidermal growth factor receptor 2 (HER2) protein is crucial for breast cancer diagnosis and targeted therapy selection.
  • Single-cell analysis presents challenges in overcoming tumor heterogeneity and improving patient outcomes.
  • Existing methods for HER2 detection may lack the sensitivity and precision required for single-cell resolution.

Purpose of the Study:

  • To develop a novel droplet microfluidic system for ultrasensitive and quantitative analysis of HER2 protein expression on single cells.
  • To enhance signal amplification for improved sensitivity and reduced fluctuation in single-cell analysis.
  • To provide a reliable platform for evaluating HER2-positive tumors and guiding personalized treatment strategies.

Main Methods:

  • A microfluidic device utilizing a tapered capillary bundle to generate 160 μm droplets.
  • An instant cation exchange signal amplification strategy involving Immuno-CdS, Ag+, Cd2+, and Rhod-5N fluorescence.
  • Quantitative analysis of HER2 protein on simulated single cells (HER2-modified PS microspheres) and SK-BR-3 cancer cells.

Main Results:

  • Achieved a limit of detection of 11.372 pg mL−1 for HER2 protein.
  • Demonstrated a 4.2-fold reduction in relative standard deviation compared to traditional immunofluorescence (2.89% vs 12.21%).
  • Successfully quantified HER2 protein on SK-BR-3 cells, ranging from 205.26 to 9862.954 pg mL−1 (2.038 × 10^5 to 9.795 × 10^6 molecules).

Conclusions:

  • The developed droplet microfluidic system coupled with instant cation exchange signal amplification enables highly sensitive and quantitative single-cell analysis of HER2 protein.
  • This innovative approach significantly improves detection sensitivity and reduces signal variability, outperforming traditional methods.
  • The platform offers a universal solution for sensitive single-cell analysis, contributing to the precise evaluation of HER2-positive tumors and advancing breast cancer diagnostics.