Quantitative chemometric phenotyping of three-dimensional liver organoids by Raman spectral imaging

Vernon LaLone; Aleksandra Aizenshtadt; John Goertz; Frøydis Sved Skottvoll; Marco Barbero Mota; Junji You; Xiaoyu Zhao; Henriette Engen Berg; Justyna Stokowiec; Minzhi Yu; Anna Schwendeman; Hanne Scholz; Steven Ray Wilson; Stefan Krauss; Molly M Stevens

doi:10.1016/j.crmeth.2023.100440

Quantitative chemometric phenotyping of three-dimensional liver organoids by Raman spectral imaging

Cell Rep Methods. 2023 Mar 31;3(4):100440. doi: 10.1016/j.crmeth.2023.100440. eCollection 2023 Apr 24.

Authors

Vernon LaLone^{1

2}, Aleksandra Aizenshtadt³, John Goertz¹, Frøydis Sved Skottvoll^{3

4}, Marco Barbero Mota¹, Junji You¹, Xiaoyu Zhao¹, Henriette Engen Berg⁴, Justyna Stokowiec³, Minzhi Yu^{5

6}, Anna Schwendeman^{5

6}, Hanne Scholz^{3

7

8}, Steven Ray Wilson^{3

4}, Stefan Krauss^{3

9}, Molly M Stevens^{1

2}

Affiliations

¹ Department of Materials, Department of Bioengineering and Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
² Hybrid Technology Hub-Centre of Excellence, Imperial College London, London SW7 2AZ, UK.
³ Hybrid Technology Hub-Centre of Excellence, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, P.O. Box 1112, Blindern, 0317 Oslo, Norway.
⁴ Department of Chemistry, University of Oslo, P.O. Box 1033, Blindern, 0315 Oslo, Norway.
⁵ Department of Pharmaceutical Sciences, University of Michigan, Ann Arbor, MI 48109, USA.
⁶ Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
⁷ Department of Transplant Medicine, Oslo University Hospital, Oslo, Norway.
⁸ Institute for Surgical Research, Oslo University Hospital, Oslo, Norway.
⁹ Department of Immunology and Transfusion Medicine, Oslo University Hospital, P.O. Box 4950, Nydalen, 0424 Oslo, Norway.

Abstract

Confocal Raman spectral imaging (RSI) enables high-content, label-free visualization of a wide range of molecules in biological specimens without sample preparation. However, reliable quantification of the deconvoluted spectra is needed. Here we develop an integrated bioanalytical methodology, qRamanomics, to qualify RSI as a tissue phantom calibrated tool for quantitative spatial chemotyping of major classes of biomolecules. Next, we apply qRamanomics to fixed 3D liver organoids generated from stem-cell-derived or primary hepatocytes to assess specimen variation and maturity. We then demonstrate the utility of qRamanomics for identifying biomolecular response signatures from a panel of liver-altering drugs, probing drug-induced compositional changes in 3D organoids followed by in situ monitoring of drug metabolism and accumulation. Quantitative chemometric phenotyping constitutes an important step in developing quantitative label-free interrogation of 3D biological specimens.

Keywords: Raman spectral imaging; drug metabolism; hepatotoxicity; liver organoids; organoid benchmarking; qRamanomics; quantitative chemotyping.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Chemometrics*
Diagnostic Imaging
Hepatocytes
Liver* / diagnostic imaging
Organoids

Abstract

Publication types

MeSH terms

Grants and funding