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Related Experiment Videos

Quantifying the geometry of micropipets.

C L Bowman1, A M Ruknudin

  • 1Department of Physiology and Biophysics, School of Medicine, State University of New York at Buffalo, and Buffalo Institute for Medical Research, VA Medical Center, 14214-3005, USA. bowman@acsu.buffalo.edu

Cell Biochemistry and Biophysics
|December 11, 1999
PubMed
Summary
This summary is machine-generated.

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Accurate micropipet tip characterization is crucial for biological membrane studies. Digital manometric method (DMM) and bubble number method (BNM) were compared, with DMM showing high accuracy comparable to scanning electron microscopy (SEM).

Area of Science:

  • Biophysics
  • Materials Science
  • Microfluidics

Background:

  • Precise internal diameter (ID) of micropipet tips is critical for studying biological membranes.
  • Tip size significantly influences the ability to analyze membrane properties.
  • Existing methods for tip characterization require evaluation for accuracy and applicability.

Purpose of the Study:

  • To compare the Digital Manometric Method (DMM) and Bubble Number Method (BNM) for micropipet tip characterization.
  • To evaluate different models (Laplace's equation and a modified form) within the DMM.
  • To assess the impact of surface properties on tip performance.

Main Methods:

  • Digital Manometric Method (DMM) using a digital manometer to measure pressure.
  • Bubble Number Method (BNM) for comparison.

Related Experiment Videos

  • Scanning Electron Microscopy (SEM) for direct measurement of micropipet tip internal diameters.
  • Testing with water and methanol to evaluate surface hydrophobicity effects.

    • Main Results:

    • Micropipet tips exhibited slight asymmetry in ID (approx. 5% difference).
    • Model I (Laplace's equation) overestimated larger IDs by 2%; Model II (modified equation) overestimated smaller IDs by 2%.
    • Uncertainties in ID measurements ranged from 20 to 100 nm for tips between 1.00 and 5.00 microm.
    • Hydrophobic surface treatment significantly reduced threshold pressures in water but not in methanol.
    • DMM demonstrated insensitivity to atmospheric pressure changes, unlike BNM.
    • DMM accuracy and precision approached that of SEM.

    Conclusions:

    • The Digital Manometric Method (DMM) provides accurate and precise measurements of micropipet tip internal diameters, comparable to SEM.
    • DMM is particularly valuable for selecting micropipets for patch-clamp studies of small vesicles (< 10 microm).
    • This method facilitates systematic micropipet selection for various experimental applications.