Automated gold particle quantification of immunogold labeled micrographs.

J Neurosci Methods

Oslo University Hospital, Department of Neurology, N-0027 Oslo, Norway; Letten Centre and GliaLab, Division of Physiology, Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, N-0317 Oslo, Norway. Electronic address:

Published: July 2017

AI Article Synopsis

Article Abstract

Background: Immunogold cytochemistry is the method of choice for precise localization of antigens on a subcellular scale. The process of immunogold quantification in electron micrographs is laborious, especially for proteins with a dense distribution pattern.

New Methods: Here I present a MATLAB based toolbox that is optimized for a typical immunogold analysis workflow. It combines automatic detection of gold particles through a multi-threshold algorithm with manual segmentation of cell membranes and regions of interests.

Results: The automated particle detection algorithm was applied to a typical immunogold dataset of neural tissue, and was able to detect particles with a high degree of precision. Without manual correction, the algorithm detected 97% of all gold particles, with merely a 0.1% false-positive rate.

Comparisons With Existing Method(s): To my knowledge, this is the first free and publicly available software custom made for immunogold analyses. The proposed particle detection method compares favorably to previously published algorithms.

Conclusions: The software presented here will be valuable tool for researchers in neuroscience working with immunogold cytochemistry.

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http://dx.doi.org/10.1016/j.jneumeth.2017.05.018DOI Listing

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