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Abstract: TH-PO571

A Novel Pipeline for the Tissue Transformation, 3D Imaging, and Visualisation in Virtual Space of Optically Cleared Human Renal Tissue

Session Information

  • Pathology and Lab Medicine
    November 03, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: Pathology and Lab Medicine

  • 1700 Pathology and Lab Medicine

Authors

  • Siew, Keith, University College London, London, London, United Kingdom
  • Li, Zhongwang, University College London, London, London, United Kingdom
  • Walsh, Stephen B., University College London, London, London, United Kingdom
Background

Kidney biopsies have been the gold standard for diagnosis of pathological kidney diseases for almost 70 years. However, due to the small volume of tissue obtained from a needle biopsy traditional histopathological examinations are limited by the number of sections that can be obtained, with no two sections capturing the exact same histological features, sometimes necessitating multiple biopsies to be taken to ensure there is enough material to work with & that an accurate representation of the kidney histology is obtained. Developing a non-destructive 3D histopathology would allow for 60+ stains to be colocalised within their intact anatomical context, therefore reducing the risk of missing pathologies & need to re-biopsy patients.

Methods

Hunan renal biopsies & tranplant rejected tissue routinely donated for research at the Royal Free Hospital were used for this study. These samples were fixed in formaldehyde, before undergoing tissue transformation following the SHIELD protocol (Park et al,. 2018. doi:10.1038/nbt.4281). These were then delipidated & stained using the LifeCavas Technolgies platforms, & optically cleared by refractive index matching prior to rapid 3D fluorescent lightsheet microscopy. These data were then preprocessed using imageJ & Imaris, before being imported into Syglass for 3D virtual reality visualisation using Oculus Quest 2 headsets.

Results

In less than 24h were able to take fresh tissue, rapidly transform, multiplex stain and image immune cell infiltration (CD3), antibodies (IgA, IgM, IgG), complement (C1q), endothelium (CD31), Interstitium (Collagen IV), Nuclei (Histone H3), cytosol (Eosin) or tubule segment markers (Tomato Lectin - DCT/TAL marker( green); see figure 1). We were able to visualise these data with a histopathologist in virtual 3D space, where it was possible toggle between the different markers, reorientate and resection the sample with ease.

Conclusion

See virtual reality headest demo at presentation.