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MIPAR and ImageJ FIJI as Tools for Electron Microscopy Quantification of Amyloid Fibrils
Charles A Garcia1, Veena Prasad1, Truman C Gamblin1
1Department of Neuroscience, Developmental, and Regenerative Biology, University of Texas at San Antonio, 1 UTSA Circle, San Antonio, Texas 78249, United States.
Abstract:
Measuring of tau filaments is an important method that can provide useful information in the study of tau in vitro. However, methods such as right-angle laser light scattering and Thioflavin T fluorescence assay only provide bulk information on the amount of tau aggregation that is occurring. Electron microscopy (EM) can be used to provide a semiquantitative method on the lengths of individual filaments and provide a length distribution of tau aggregates. The issue with quantifying tau aggregation through EM is that it can be time costly if done manually. Here we explore two different programs, MIPAR and ImageJ FIJI, as methods to automate the quantification of EM grids. Using both programs to measure filaments produced from inducing 2N4R tau with the fatty acid arachidonic acid (ARA), we are able to reliably measure filaments producing similar results from MIPAR and ImageJ, with these methods applicable to other filamentous biological structures.
Insights
Automating the measurement of tau filaments using electron microscopy (EM) with MIPAR and ImageJ FIJI software offers a reliable and efficient alternative to manual quantification. These methods accurately assess tau aggregate lengths and distributions, crucial for in vitro tau studies.
Area of Science:
- Biochemistry
- Microscopy
- Computational Biology
Background:
- In vitro studies of tau filaments are essential for understanding tauopathies.
- Current methods like Thioflavin T fluorescence and laser light scattering provide only bulk measurements of tau aggregation.
- Electron microscopy (EM) offers detailed filament length information but manual quantification is time-consuming.
Purpose of the Study:
- To explore automated quantification of EM grids for tau filaments using MIPAR and ImageJ FIJI.
- To compare the reliability and efficiency of MIPAR and ImageJ FIJI for measuring tau aggregate lengths and distributions.
- To establish applicable methods for other filamentous biological structures.
Main Methods:
- Induction of 2N4R tau filaments using arachidonic acid (ARA).
- Acquisition of EM images of tau aggregates.
- Automated quantification of filament lengths and distributions using MIPAR and ImageJ FIJI software.
Main Results:
- Both MIPAR and ImageJ FIJI programs reliably quantified tau filaments.
- The automated methods produced similar and consistent results for filament measurements.
- The developed quantification approaches are applicable to other filamentous biological structures.
Conclusions:
- Automating EM grid analysis with MIPAR and ImageJ FIJI significantly improves the efficiency of tau filament quantification.
- These validated methods provide accurate semiquantitative data on tau aggregate length distributions.
- The approach is versatile and can be applied to the study of various filamentous biological structures.
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