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Related Concept Videos

Patch Clamp01:18

Patch Clamp

7.3K
Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...
7.3K

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Related Experiment Video

Updated: Mar 19, 2026

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
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A Single-Cell-Type Real-Time PCR Method Based on a Modified Patch-Pipette Cell Harvesting System.

Yuanlong Song1,2, Miaomiao Zhang1, Xiaoqing Tao1

  • 1Department of Physiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Rd., Wuhan, China.

Molecular Biotechnology
|June 9, 2016
PubMed
Summary

This study introduces a novel method combining single-cell and tissue-level PCR for accurate nucleic acid quantification. It overcomes limitations of both techniques by using a modified patch-clamp pipette to harvest multiple pure cells for analysis.

Keywords:
Cell harvestingMethodsPCRPatch-clampSingle-cell

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biotechnology

Background:

  • Real-time PCR (RT-PCR) is vital for nucleic acid quantification but tissue-level analysis suffers from cell mixture impurities.
  • Single-cell PCR offers purity but faces challenges in cell harvesting and processing minute nucleic acid quantities.

Purpose of the Study:

  • To develop a refined PCR method that integrates the cell purity of single-cell analysis with the ease of tissue-level analysis.
  • To enable reliable nucleic acid quantification from specific, identified cell populations.

Main Methods:

  • A modified patch-clamp pipette was utilized for harvesting multiple, selected cells of the same type.
  • This approach facilitates the processing of larger nucleic acid quantities compared to traditional single-cell methods.

Main Results:

  • The developed method successfully combines the advantages of high cell purity and increased nucleic acid yield.
  • It addresses the limitations inherent in both conventional tissue-level and single-cell PCR techniques.

Conclusions:

  • This technique is particularly advantageous for analyzing morphologically or histologically identifiable cells, such as sensory neurons or muscle fibers.
  • It offers a more precise and reliable approach to nucleic acid expression quantification in specific cell types.