Multiplex Immunofluorescent Batch Labeling of Marmoset Brain Sections

Abstract

Purpose

The common marmoset is a small nonhuman primate that has emerged as a valuable animal model in neuroscience research. Accurate analysis of brain tissue is crucial to understand marmoset neurophysiology and to model neurodegenerative diseases. Many studies to date have complemented magnetic resonance imaging (MRI) with histochemical staining rather than immunofluorescent labeling, which can generate more informative and higher resolution images. There is a need for high-throughput immunolabeling and imaging methodologies to generate resources for the burgeoning marmoset field, particularly brain histology atlases to display the organization of different cell types and other structures.

Methods and Findings

Here, we have characterized a set of marmoset-compatible fluorescent dyes and antibodies that label myelin, axons, dendrites, and the iron-storage protein ferritin, and developed a batch-style multiplex immunohistochemistry protocol to uniformly process large numbers of tissue slides for multiple cell-type specific markers.

Conclusion

We provide a practical guide for researchers interested in harnessing the potential of marmoset models to advance understanding of brain structure, function, and pathophysiology.