High-throughput 3D whole-brain quantitative histopathology in rodents.

Nicolas Souedet,Elmahdi Sadouni,Jocelyne Schulz,Dominique Briet,Thierry Delzescaux,Michel E. Vandenberghe,Anne-Sophie Hérard,Marc Dhenain,Denis Carré,Laurent Pradier,Mathieu D. Santin,Philippe Hantraye,Thomas Debeir,Pierre-Etienne Chabrier,Thomas Rooney,Véronique Blanchard

doi:10.1038/srep20958

Abstract

Histology is the gold standard to unveil microscopic brain structures and pathological alterations in humans and animal models of disease. However, due to tedious manual interventions, quantification of histopathological markers is classically performed on a few tissue sections, thus restricting measurements to limited portions of the brain. Recently developed 3D microscopic imaging techniques have allowed in-depth study of neuroanatomy. However, quantitative methods are still lacking for whole-brain analysis of cellular and pathological markers. Here, we propose a ready-to-use, automated, and scalable method to thoroughly quantify histopathological markers in 3D in rodent whole brains. It relies on block-face photography, serial histology and 3D-HAPi (Three Dimensional Histology Analysis Pipeline), an open source image analysis software. We illustrate our method in studies involving mouse models of Alzheimer’s disease and show that it can be broadly applied to characterize animal models of brain diseases, to evaluate therapeutic interventions, to anatomically correlate cellular and pathological markers throughout the entire brain and to validate in vivo imaging techniques.

Highlights

Histology is the gold standard to unveil microscopic brain structures and pathological alterations in humans and animal models of disease
We propose an integrated approach that relies on block-face photography[25,26], serial histology and 3D-HAPi (3D Histology Analysis Pipeline), an innovative image analysis pipeline freely available for non-commercial use
We evaluated the effect of a new anti-amyloid immunotherapy in 8-months-old APP/PS1 transgenic mice in the entire brain and in four regions of interest (ROIs)

Summary

Introduction

Histology is the gold standard to unveil microscopic brain structures and pathological alterations in humans and animal models of disease. A pending question is how to make sense of this large amount of data to provide new biological insight To address these issues, we propose an integrated approach that relies on block-face photography[25,26], serial histology and 3D-HAPi (3D Histology Analysis Pipeline), an innovative image analysis pipeline freely available for non-commercial use (http://brainvisa.info). As a proof of concept of our method usability, we studied well-known mouse models of cerebral Aβ peptide deposition which is a neuropathological hallmark of Alzheimer’s disease[27] In these mice, we thoroughly characterized Aβ peptide deposition, we evaluated the brain-wide effect of a new anti-amyloid immunotherapy, we spatially correlated neuroinflammatory cell markers with Aβ peptide deposition across the whole brain and we registered 3D histopathological data with in vivo contrast-enhanced MRI

Methods

Results

Conclusion