An Optimized Freeze‐Dry Multimodal Workflow for Sequential Micro‐CT Imaging, Histology, and Molecular Profiling: A Use Case in Human Liver Fibrosis
9 affiliations
- 1Diagnostic and Research Institute of Pathology Medical University of Graz Graz Austria
- 2Institute of Biomedical Sciences, Faculty of Medicine University of Maribor Maribor Slovenia
- 3Division of Gastroenterology and Hepatology Department of Internal Medicine Medical University of Graz Graz Austria
- 4Research Unit for Translational Nuclear Receptor Research Graz Austria
- 5BioTechMed Graz Graz Austria
- 6ZLP—Zentrum für Leber‐ und Pankreaspathologie, ADK Diagnostics Vienna Austria
- 7Clinical Institute of Medical and Chemical Laboratory Diagnostics Medical University of Graz Graz Austria
- 8Department of Pathology, Queen Elizabeth II Health Sciences Centre Dalhousie University Halifax Nova Scotia Canada
- 9University of Maribor Faculty of Medicine Department of Pharmacology Maribor Slovenia
Abstract
Classical histology remains the gold standard for liver fibrosis assessment but is limited by destructive sectioning, 2D information, and sampling bias. Micro‐CT offers non‐destructive 3D imaging, yet conventional contrast agents can compromise tissue integrity and downstream analyses. This study establishes a resource‐efficient multimodal workflow that enables high‐quality imaging and subsequent histological and molecular testing from the same specimen. FFPE normal and cirrhotic liver tissues (n = 3 each) undergo an optimized micro‐CT protocol: DPBS wash, 30% eosin incubation, dehydration, snap‐freezing, and lyophilization. After imaging, tissues are rehydrated, re‐embedded in paraffin, and processed for routine histology, immunohistochemistry, RNA extraction with qPCR, and targeted NGS (800‐gene panel). The protocol yields high‐contrast, artifact‐free micro‐CT images with clear visualization of nuclei, sinusoidal networks, portal structures, and fibrotic septa (∼20x–40x optical equivalent). Volumetrics show minimal fibrosis in normal liver (2.8%) versus extensive remodeling in cirrhosis (43.5%). Morphology and antigenicity are preserved (H&E, PSR, IHC). RNA quality supports qPCR, revealing upregulation of inflammatory and cirrhosis‐related genes in cirrhosis. Targeted NGS libraries are successfully generated, with protocol‐dependent performance differences. This minimally destructive workflow maximizes information yield from limited tissue, enabling integrated imaging and molecular profiling with broad utility in pathology, biobanking, and translational research.
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BibTeX
@article{skok2026optimized,
title = {{An Optimized Freeze‐Dry Multimodal Workflow for Sequential Micro‐CT Imaging, Histology, and Molecular Profiling: A Use Case in Human Liver Fibrosis}},
author = {Kristijan Skok and Laura Činč Ćurić and Jernej Vajda and Silvia Schauer and Martin Wagner and Rudolf Stauber and Carolin Lackner and Karl Kashofer and Markus Plass and Giovanny Rodriguez Blanco and Ian R. Wanless and Uroš Maver and Katrin Panzitt and Boštjan Vihar},
journal = {Advanced Science},
year = {2026},
month = aug,
doi = {10.1002/advs.77406},
url = {https://doi.org/10.1002/advs.77406},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - An Optimized Freeze‐Dry Multimodal Workflow for Sequential Micro‐CT Imaging, Histology, and Molecular Profiling: A Use Case in Human Liver Fibrosis
AU - Kristijan Skok
AU - Laura Činč Ćurić
AU - Jernej Vajda
AU - Silvia Schauer
AU - Martin Wagner
AU - Rudolf Stauber
AU - Carolin Lackner
AU - Karl Kashofer
AU - Markus Plass
AU - Giovanny Rodriguez Blanco
AU - Ian R. Wanless
AU - Uroš Maver
AU - Katrin Panzitt
AU - Boštjan Vihar
PY - 2026
DA - 2026/08/28
JO - Advanced Science
DO - 10.1002/advs.77406
UR - https://doi.org/10.1002/advs.77406
AB - Classical histology remains the gold standard for liver fibrosis assessment but is limited by destructive sectioning, 2D information, and sampling bias. Micro‐CT offers non‐destructive 3D imaging, yet conventional contrast agents can compromise tissue integrity and downstream analyses. This study establishes a resource‐efficient multimodal workflow that enables high‐quality imaging and subsequent histological and molecular testing from the same specimen. FFPE normal and cirrhotic liver tissues (n = 3 each) undergo an optimized micro‐CT protocol: DPBS wash, 30% eosin incubation, dehydration, snap‐freezing, and lyophilization. After imaging, tissues are rehydrated, re‐embedded in paraffin, and processed for routine histology, immunohistochemistry, RNA extraction with qPCR, and targeted NGS (800‐gene panel). The protocol yields high‐contrast, artifact‐free micro‐CT images with clear visualization of nuclei, sinusoidal networks, portal structures, and fibrotic septa (∼20x–40x optical equivalent). Volumetrics show minimal fibrosis in normal liver (2.8%) versus extensive remodeling in cirrhosis (43.5%). Morphology and antigenicity are preserved (H&E, PSR, IHC). RNA quality supports qPCR, revealing upregulation of inflammatory and cirrhosis‐related genes in cirrhosis. Targeted NGS libraries are successfully generated, with protocol‐dependent performance differences. This minimally destructive workflow maximizes information yield from limited tissue, enabling integrated imaging and molecular profiling with broad utility in pathology, biobanking, and translational research.
SN - 2198-3844
ER -