Multimodal imaging of a liver-on-a-chip model using labelled and label-free optical microscopy techniques †

Majer, Jan, Alex, Aneesh, Shi, Jindou, Chaney, Eric J., Mukherjee, Prabuddha, Spillman, Darold R., Marjanovic, Marina, Newman, Carla F., Groseclose, Reid M., Watson, Peter D. ORCID: https://orcid.org/0000-0003-0250-7852, Boppart, Stephen A. and Hood, Steve R. 2024. Multimodal imaging of a liver-on-a-chip model using labelled and label-free optical microscopy techniques †. Lab on a Chip 24 (19) , pp. 4594-4608. 10.1039/d4lc00504j

PDF - Published Version Download (6MB)

Abstract

A liver-on-a-chip model is an advanced complex in vitro model (CIVM) that incorporates different cell types and extracellular matrix to mimic the microenvironment of the human liver in a laboratory setting. Given the heterogenous and complex nature of liver-on-a-chip models, brightfield and fluorescence-based imaging techniques are widely utilized for assessing the changes occurring in these models with different treatment and environmental conditions. However, the utilization of optical microscopy techniques for structural and functional evaluation of the liver CIVMs have been limited by the reduced light penetration depth and lack of 3D information obtained using these imaging techniques. In this study, the potential of both labelled as well as label-free multimodal optical imaging techniques for visualization and characterization of the cellular and sub-cellular features of a liver-on-a-chip model was investigated. (1) Cellular uptake and distribution of Alexa 488 (A488)-labelled non-targeted and targeted antisense oligonucleotides (ASO and ASO-GalNAc) in the liver-on-a-chip model was determined using multiphoton microscopy. (2) Hyperspectral stimulated Raman scattering (SRS) microscopy of the C–H region was used to determine the heterogeneity of chemical composition of circular and cuboidal hepatocytes in the liver-on-a-chip model in a label-free manner. Additionally, the spatial overlap between the intracellular localization of ASO and lipid droplets was explored using simultaneous hyperspectral SRS and fluorescence microscopy. (3) The capability of light sheet fluorescence microscopy (LSFM) for full-depth 3D visualization of sub-cellular distribution of A488-ASO and cellular phenotypes in the liver-on-a-chip model was demonstrated. In summary, multimodal optical microscopy is a promising platform that can be utilized for visualization and quantification of 3D cellular organization, drug distribution and functional changes occurring in liver-on-a-chip models, and can provide valuable insights into liver biology and drug uptake mechanisms by enabling better characterization of these liver models.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Additional Information:	License information from Publisher: LICENSE 1: URL: https://creativecommons.org/licenses/by/3.0/, Start Date: 2024-09-11
Publisher:	Royal Society of Chemistry
ISSN:	1473-0197
Date of First Compliant Deposit:	12 September 2024
Date of Acceptance:	26 August 2024
Last Modified:	12 Nov 2024 13:59
URI:	https://orca.cardiff.ac.uk/id/eprint/172054

Actions (repository staff only)

Edit Item