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A simple method for measuring organotypic tissue slice culture thickness.

Yifat Guy1, Amy E Rupert, Mats Sandberg

  • 1Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA. yig1@pitt.edu

Journal of Neuroscience Methods
|April 19, 2011
PubMed
Summary

This study introduces a quick ohmmeter method to measure tissue slice thickness without harming cultures. This technique accurately tracks changes in slice thickness during different culture conditions.

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Area of Science:

  • Neuroscience
  • Biotechnology
  • Tissue Engineering

Background:

  • Accurate measurement of tissue slice thickness is crucial for understanding cellular behavior and experimental outcomes.
  • Existing methods for measuring slice thickness, such as optical measurements, often require sacrificing the tissue sample.
  • There is a need for a non-invasive, rapid, and accurate method to determine tissue slice thickness in real-time.

Purpose of the Study:

  • To present a novel, simple, and non-invasive method for measuring tissue slice thickness using an ohmmeter.
  • To evaluate the method's accuracy and compare it with established techniques.
  • To assess the impact of different culturing protocols and conditions on tissue slice thickness.

Main Methods:

  • A circuit was designed comprising a metal probe, an ohmmeter, a counter electrode, culture medium, and the tissue slice.

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  • The probe and electrode were positioned on opposite sides of the tissue slice.
  • Thickness was determined by measuring the probe's position change upon contact with the tissue surface versus the supporting membrane.

    • Main Results:

    • The ohmmeter method provided accurate thickness measurements without compromising slice viability.
    • Tissue slice thickness showed an initial decrease in the first 4 days of culture, stabilizing thereafter.
    • Culture medium type and initial explant thickness were identified as factors influencing final slice thickness.

    Conclusions:

    • The proposed ohmmeter-based technique offers a straightforward, rapid (approx. 1 minute per culture), and non-destructive approach for measuring tissue slice thickness.
    • This method facilitates longitudinal studies of tissue slice cultures by enabling repeated measurements.
    • The findings contribute to improved methodologies in tissue engineering and neuroscience research, particularly for organotypic slice cultures.