Uncovering microstructural architecture from histology

Microstructural tissue organization underlies the complex connectivity of the brain and controls the properties of connective, muscle, and epithelial tissue. However, discerning microstructural architecture with high resolution for large fields of view remains prohibitive. We address this challenge with computational scattered light imaging (ComSLI), which exploits the anisotropic light scattering of aligned structures. Using a rotating light source and a high-resolution camera, ComSLI determines fiber architecture with micrometer resolution from histological sections across preparation and staining protocols. We show complex fiber architecture in brain and non-brain sections, including histological paraffin-embedded sections with various stains, and demonstrate its applicability on animal and human tissue, including disease cases with altered microstructure. ComSLI opens new avenues for investigating fiber architecture in new and archived sections across organisms, tissues, and disease..., Sample preparation
Whole human brain (BigBrain) sections
Two human brains, obtained within 24 hours after death and without neurological disorders, were fixed in 4% formaldehyde, dehydrated in increasing alcohol series (80%, 90%, 96%, 100% ethanol for at least one week each), embedded in a solvent (chloroform) for one week, and embedded in 57-60°C paraffin solution for two months. After hardening, the brains were coronally cut into 20 µm-thin sections from anterior to posterior with a large-scale microtome (Leica SM2500 Microtome) and mounted on a 37°C heat plate in gelatin solution. The sections were placed in a decreasing alcohol series to remove the paraffin and then placed in a staining solution to highlight neuronal cell bodies: the sections of the first brain (30 years old, male) were stained with silver following the protocol of Merker (53); the sections of the second brain (71 years old, male) were stained with Cresyl-violet. After staining, the sections were again dehydrated in..., , # Uncovering **microstructural** architecture from histology

[https://doi.org/10.5061/dryad.02v6wwqb2](https://doi.org/10.5061/dryad.02v6wwqb2)

This dataset includes all raw figures of the data presented in the manuscript \"*Uncovering microstructural* *architecture from Histology*\".

## Description of the data and file structure

Each file is named under the Figure number and panel in which it is presented in the manuscript.

All brain image files are in .tiff format, except the SupplementaryMovie1.gif.