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Updated: May 27, 2026
Polarization-Sensitive Two-Photon Microscopy for a Label-Free Amyloid Structural Characterization
Published on: September 8, 2023
Semi-automated methods for simulation and measurement of amyloid fiber distributions obtained from transmission
1Institute of Basic Medical Science, University of Tsukuba, Tsukuba-shi, Ibaraki-ken 305-8577, Japan. damienhall@md.tsukuba.ac.jp
Abstract:
Transmission electron microscopy (TEM) is the standard procedure for qualitatively confirming the presence of amyloid fibers in a protein aggregation reaction product. However, extracting quantitative information about the amyloid size distribution from the electron micrographs is a nontrivial problem. Here we describe methods for (i) the simulation of pseudo-TEM images of amyloid fiber distributions having known characteristic properties and (ii) the semi-automated processing of experimental TEM images of amyloid fibers to produce two-dimensional histogram plots reflecting either the distribution of amyloid length and width or, alternatively, the distribution of width and fiber rigidity/persistence. The processing method is fully automatic when the density of fibers on the grid is sufficiently low (such that the adsorbed fibers do not touch) and is semi-automatic (requiring some user decision making) when the fibers are overlapping. Termed "ADM" (for Amyloid Distribution Measurement), the program suite is written in MATLAB code and is available on request from the author.
Insights
This study introduces the Amyloid Distribution Measurement (ADM) software for quantifying amyloid fiber size and properties from transmission electron microscopy (TEM) images. ADM provides automated analysis for low-density samples and semi-automated for high-density, enabling precise amyloid characterization.
Area of Science:
- Biophysics
- Materials Science
- Biochemistry
Background:
- Transmission electron microscopy (TEM) is crucial for identifying amyloid fibers in protein aggregates.
- Quantitative analysis of amyloid fiber size distribution from TEM images presents significant challenges.
Purpose of the Study:
- To develop methods for simulating pseudo-TEM images of amyloid distributions.
- To create a semi-automated software tool for quantitative analysis of experimental TEM images of amyloid fibers.
Main Methods:
- Simulation of pseudo-TEM images with defined amyloid properties.
- Development of the Amyloid Distribution Measurement (ADM) software using MATLAB.
- Implementation of automated and semi-automated image processing for fiber analysis.
Main Results:
- The ADM software generates 2D histogram plots of amyloid length/width or width/rigidity.
- Automated processing is effective for low-density fiber samples.
- Semi-automated processing accommodates overlapping fibers in high-density samples.
Conclusions:
- The ADM software provides a robust solution for quantitative amyloid fiber analysis from TEM data.
- This tool enhances the characterization of amyloid aggregation processes.
- ADM facilitates precise measurement of amyloid fiber dimensions and properties.
