3D reconstruction and spatial quantitative analysis of NAFLD

Introduction:

Non-alcoholic fatty liver disease (NAFLD) is the most common form of chronic liver disease in Western countries. Classical 2D histological approaches are limited and cannot provide the proper description of the morphological changes occurring during disease progression. We have developed high-resolution imaging and quantitative multi-parametric spatial image analysis approaches aimed to identify structural changes in cell and tissue organization associated with NAFLD.

Material and methods:

To cover the disease progression, human samples from different disease stages were collected (healthy control, steatosis and non-alcoholic steatohepatitis (NASH)). Liver sections (100 µm thick) were stained for cell border, bile canaliculi, sinusoids, lipid droplets and nucleus, optically cleared and imaged using multiphoton microscopy. The Motion Tracking software (Morales-Navarrete et al., eLIFE 2015) was used for segmentation and quantitative analysis of the acquired images.

Results:

We have established a protocol to perform 3D reconstruction and spatial quantitative analysis from various disease stages of NAFLD in human liver samples. We provide a quantitative description of the bile canaliculi and sinusoidal networks from central to portal vein. We can also extract precise quantitative morphometric parameters of liver tissue focusing on hepatocyte cells (e.g. hepatocyte ploidy, volume, surface area, apical, lateral and basal surface, number of neighbors, lipid droplets).

Perspective/Conclusion:

This method will allow exploring new aspects of non-alcoholic fatty liver disease that have not been described before due to the limiting 2D histological approach, thus contributing to the understanding of NAFLD liver physiopathology.