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A practical guide to working with nanopipettes.

Dominik Duleba1, Adria Martínez-Aviñó1, Robert P Johnson1

  • 1School of Chemistry, University College Dublin, Belfield, Ireland. robert.johnson@ucd.ie.

The Analyst
|December 5, 2025
PubMed
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This guide offers practical solutions for nanopipette fabrication, filling, and surface modification challenges. It aims to improve reproducibility and throughput for nanopore research, lowering the entry barrier for new labs.

Area of Science:

  • Materials Science
  • Analytical Chemistry
  • Biophysics

Background:

  • Nanopipettes are versatile solid-state nanopore platforms for single-particle analysis and high-sensitivity measurements.
  • Despite demonstrated capabilities, practical challenges in fabrication, filling, and surface modification limit their widespread adoption and reproducibility.
  • Low throughput and high entry barriers hinder progress in nanopipette research.

Purpose of the Study:

  • To provide a practical guide addressing common technical hurdles in nanopipette research.
  • To share accumulated practical experience for overcoming fabrication, filling, and surface modification challenges.
  • To facilitate a holistic workflow for enhanced throughput and reproducibility in nanopipette applications.

Main Methods:

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  • Detailed fabrication protocols emphasizing environmental control and instrument maintenance for reproducibility.
  • Analysis of nanopipette geometries to optimize filling and surface modification.
  • Exploration of filling procedures, considering surface wetting, capillary forces, and surface chemistry.
  • Discussion of silanization techniques, particularly vapor-phase silanization, for interior surface modification.
  • Principles for surface grafting reactions to maximize throughput and reproducibility.

    • Main Results:

    • Established protocols for reproducible nanopipette fabrication through controlled environmental conditions and instrument upkeep.
    • Identified optimal nanopipette geometries that simplify filling and surface modification processes.
    • Provided insights into mitigating filling issues by understanding surface wetting, capillary forces, and chemistry.
    • Demonstrated effective vapor-phase silanization for consistent nanopipette interior surface modification.
    • Outlined design principles for surface grafting, enhancing throughput and reproducibility.

    Conclusions:

    • Addressing interconnected challenges in nanopipette geometry, filling, and surface modification is crucial.
    • A holistic workflow, considering each stage in tandem, significantly improves research reproducibility and throughput.
    • This practical guide aims to lower the entry barrier, enabling wider adoption of nanopipette technology.