Visible and Near-Infrared hyperspectral imaging (HSI) can reliably quantify CD3 and CD45 positive inflammatory cells in myocarditis: Pilot study on formalin-fixed paraffin-embedded specimens from myocard obtained during autopsy

[Display omitted] •HSI can reliably count immunohistochemically stained inflammatory cells in suspected myocardial disease.•A high correlation was present between the results obtained by VA compared to HSI and DIA for both markers, CD3 and CD45.•HSI can be expanded e.g. towards analysis of multiple stainings or unstained slides.•HSI can be a reliable, cost-effective and quick alternative for analysis of pathological specimens. To implement Hyperspectral Imaging (HSI) as a tool for quantifying inflammatory cells in tissue specimens by the example of myocarditis in a collective of forensic patients. 44 consecutive patients with suspected myocardial inflammation at autopsy, diagnosed between 2013 and 2018 at the Institute of ForensicMedicine, Medical University of Innsbruck, were selected for this study. Using the IMEC SNAPSCAN camera, visible and near infrared hyperspectral images were collected from slides stained with CD3 and CD45 to assess quantity and spatial distribution of positive cells. Results were compared with visual assessment (VA) and conventional digital image analysis (DIA). Finally, specimens of 40 patients were evaluated, of whom 36 patients (90%) suffered from myocarditis, two patients (5%) had suspected healing/healed myocarditis, and two did no have myocarditis (5%). The amount of CD3 and CD45 positive cells did not differ significantly between VA, HSI, and DIA (pVA/HSI/DIA = 0.46 for CD3 and 0.81 for CD45). Coheńs Kappa showed a very high correlation between VA versus HSI, VA versus DIA, and HSI versus DIA for CD3 (Coheńs Kappa = 0.91, 1.00, and 0.91, respectively). For CD45 an almost as high correlation was seen for VA versus HSI and HSI versus DIA (Coheńs Kappa = 0.75 and 0.70) and VA versus DIA (Coheńs Kappa = 0.89). HSI is a reliable and objective method to count inflammatory cells in tissue slides of suspected myocarditis. Implementation of HSI in digital pathology might further expand the possibility of a sophisticated method.