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Related Experiment Videos

Using atomic force microscopy to investigate patch-clamped nuclear membrane

T Danker1, M Mazzanti, R Tonini

  • 1Physiologisches Institut, Würzburg, Germany.

Cell Biology International
|October 13, 1998
PubMed
Summary
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Nuclear patch clamp studies electrical activity of the nuclear envelope (NE). Combining patch clamp with atomic force microscopy (AFM) reveals NE structure and identifies contaminants like endoplasmic reticulum.

Area of Science:

  • Cell Biology
  • Biophysics
  • Molecular Biology

Background:

  • Nuclear patch clamp is an emerging technique to study the nuclear envelope's (NE) electrical properties.
  • The NE regulates macromolecular transport, acts as an ion barrier, and stores intracellular calcium.
  • Current patch clamp data on NE currents are lower than expected, suggesting a gap in understanding nuclear pore complex (NPC) function.

Purpose of the Study:

  • To investigate the structure-function relationship of the NE using a combined patch clamp and atomic force microscopy (AFM) approach.
  • To correlate electrical recordings from the NE with its structural characteristics.
  • To clarify the role of NPCs in NE electrical activity.

Main Methods:

  • Combined patch clamp electrophysiology with atomic force microscopy (AFM).

Related Experiment Videos

  • Excised NE membrane patches were electrically characterized and then transferred to a substrate for AFM imaging.
  • AFM provided high-resolution structural analysis (3 nm lateral, 0.1 nm vertical) of NE patches.

    • Main Results:

    • AFM successfully identified excised native NE patches, including NPCs, after electrical evaluation.
    • The study demonstrated that complete NEs with NPCs can be excised following functional patch clamp identification.
    • A significant finding was the identification of endoplasmic reticulum membrane contamination in patch pipettes during nuclear patch clamp experiments.

    Conclusions:

    • The combination of patch clamp and AFM is effective for analyzing NE structure-function relationships.
    • The presence of endoplasmic reticulum membranes must be carefully considered to avoid misinterpretation of nuclear patch clamp data.
    • This integrated approach advances the understanding of NE electrical properties and NPC function.