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Practicable methods for histological section thickness measurement in quantitative stereological analyses.

Cyrill Matenaers1, Bastian Popper2, Alexandra Rieger1

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Accurate histological section thickness is crucial for stereological analysis. Spectral reflectance (SR) offers a fast, precise alternative to the laborious orthogonal re-embedding (ORE) method, with less than 1% deviation.

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Area of Science:

  • Histology
  • Stereology
  • Materials Science

Background:

  • Accurate histological section thickness is vital for quantitative stereological analysis, like the physical disector method.
  • Conventional orthogonal re-embedding (ORE) is time-consuming and prone to errors from non-orthogonal positioning.

Purpose of the Study:

  • To present an improved ORE method for accurate section thickness measurement, correcting for oblique sectioning.
  • To evaluate spectral reflectance (SR) as a fast and accurate alternative for determining plastic section thicknesses.

Main Methods:

  • An improved ORE method involving a calibration foil to determine section angle and correct thickness measurements.
  • Evaluation of spectral reflectance (SR) using a commercial reflectometer to measure thicknesses of Epon and GMA/MMA plastic sections.

Main Results:

  • The improved ORE method allows for correction of section thickness values for oblique sectioning.
  • Spectral reflectance (SR) precisely measured section thicknesses (0.5-3 μm) within seconds.
  • SR measurements showed less than 1% deviation compared to ORE.

Conclusions:

  • Spectral reflectance (SR) is a practicable, fast, and accurate method for determining plastic section thicknesses.
  • SR provides reliable section thickness measurements essential for accurate quantitative stereological analysis.