High-Resolution Imaging of Tumor Spheroids and Organoids Enabled by Expansion Microscopy

Frontiers in Molecular Biosciences

Kyra Kuhnigk¹  Valentina Carannante²  Henrik Ring³  Tatsiana Tararuk³  Finn Hallböök³  Hjalmar Brismar⁴  Hans Blom⁴  Björn Önfelt⁴  Steven J. Edwards⁴ 

[1] Center for Hematology and Regenerative Medicine, Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden;Department of Microbiology, Tumor and Cell Biology, Science for Life Laboratory, Karolinska Institutet, Stockholm, Sweden;Department of Neuroscience, BMC, Uppsala University, Uppsala, Sweden;Science for Life Laboratory, Department of Applied Physics, KTH Royal Institute of Technology, Stockholm, Sweden;

关键词: expansion;    microscopy;    spheroid;    organoid;    lightsheet;    imaging;   

DOI  :  10.3389/fmolb.2020.00208

来源: DOAJ

【 摘 要 】

Three-dimensional cell cultures are able to better mimic the physiology and cellular environments found in tissues in vivo compared to cells grown in two dimensions. In order to study the structure and function of cells in 3-D cultures, light microscopy is frequently used. The preparation of 3-D cell cultures for light microscopy is often destructive, including physical sectioning of the samples, which can result in the loss of 3-D information. In order to probe the structure of 3-D cell cultures at high resolution, we have explored the use of expansion microscopy and compared it to a simple immersion clearing protocol. We provide a practical method for the study of spheroids, organoids and tumor-infiltrating immune cells at high resolution without the loss of spatial organization. Expanded samples are highly transparent, enabling high-resolution imaging over extended volumes by significantly reducing light scatter and absorption. In addition, the hydrogel-like nature of expanded samples enables homogenous antibody labeling of dense epitopes throughout the sample volume. The improved labeling and image quality achieved in expanded samples revealed details in the center of the organoid which were previously only observable following serial sectioning. In comparison to chemically cleared spheroids, the improved signal-to-background ratio of expanded samples greatly improved subsequent methods for image segmentation and analysis.

【 授权许可】

Unknown