Standardised TruAI Automated Quantification of Intracellular Neuromelanin Granules in Human Brain Tissue Sections

Abstract

Artificial intelligence automation; Colour identification; Computer‐assisted quantitation

Automatización de la inteligencia artificial; Identificación de colores; Cuantificación asistida por ordenador

Automatització de la intel·ligència artificial; Identificació de colors; Quantificació assistida per ordinador

Aims To standardise and automate the quantitation of human-unique neuromelanin granules in catecholamine neurons in post-mortem tissue sections from healthy individuals at different ages to understand any changes in these granules with age. Methods Five- to 6-μm-thick fixed and paraffin-embedded transverse midbrain tissue sections were supplied from 47 cases from three brain banks following ethical approvals. Sections were prepared and automated digital images acquired. Standardisation and automation of the quantification of neuromelanin granules was performed using the TruAI feature of the Olympus VS200 desktop platform. Comparisons between stained and unstained sections as well as correlations with age were performed. Results The automated platform reliably identified both stained and unstained intracellular neuromelanin granules and extracellular pigments, showing high reproducibility in measurements across laboratories using different tissue processing methods. Extraneuronal pigments were significantly smaller than intracellular neuromelanin granules. Sections processed for haematoxylin and eosin staining impacted the size and colour of both neuromelanin and the neurons containing neuromelanin. Haematoxylin made neuromelanin bluer, and the increased tissue processing made the intracellular area occupied by neuromelanin smaller in younger people. There was an increase in neuromelanin optical density and colour change (browner) with age. Conclusions The TruAI automated platform reliably quantifies individual neuromelanin granules in catecholamine neurons. Extraneuronal pigments are considerably smaller in size than intracellular neuromelanin, and intracellular neuromelanin changes its properties with age. The darkening and colour change of intracellular neuromelanin suggest an increase in eumelanin over time in healthy individuals. These changes can be reliably identified using the automated platform.

This work was supported in part by Aligning Science Across Parkinson's [020505] through the Michael J. Fox Foundation for Parkinson's Research (MJFF) and also by funding from the National Health and Medical Research Council of Australia to GMH [Investigator Grant 1176607]. For open access, the authors have applied a CC BY public copyright license to all Author Accepted Manuscripts arising from this submission.