MIPAR and ImageJ FIJI as Tools for Electron Microscopy Quantification of Amyloid Fibrils.

IF 3 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemistry Biochemistry Pub Date : 2025-05-06 Epub Date: 2025-04-08 DOI:10.1021/acs.biochem.5c00031
Charles A Garcia, Veena Prasad, Truman C Gamblin
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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.

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MIPAR和ImageJ FIJI作为电子显微镜定量淀粉样蛋白原纤维的工具。
tau蛋白细丝的测量是一种重要的方法,可以为tau蛋白的体外研究提供有用的信息。然而,直角激光散射和硫黄素T荧光测定等方法只能提供正在发生的tau聚集量的大量信息。电子显微镜(EM)可用于提供对单个细丝长度的半定量方法,并提供tau聚集体的长度分布。通过EM量化tau聚集的问题是,如果手工完成,可能会耗费时间。在这里,我们探索了两个不同的程序,MIPAR和ImageJ FIJI,作为EM网格自动化量化的方法。使用这两个程序来测量由脂肪酸花生四烯酸(ARA)诱导2N4R tau产生的细丝,我们能够可靠地测量从MIPAR和ImageJ产生类似结果的细丝,这些方法适用于其他丝状生物结构。
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来源期刊
Biochemistry Biochemistry
Biochemistry Biochemistry 生物-生化与分子生物学
CiteScore
5.50
自引率
3.40%
发文量
336
审稿时长
1-2 weeks
期刊介绍: Biochemistry provides an international forum for publishing exceptional, rigorous, high-impact research across all of biological chemistry. This broad scope includes studies on the chemical, physical, mechanistic, and/or structural basis of biological or cell function, and encompasses the fields of chemical biology, synthetic biology, disease biology, cell biology, nucleic acid biology, neuroscience, structural biology, and biophysics. In addition to traditional Research Articles, Biochemistry also publishes Communications, Viewpoints, and Perspectives, as well as From the Bench articles that report new methods of particular interest to the biological chemistry community.
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