Organisms are three dimensional and the spatial organisation of cells in a tissue is essential for its proper function. Classical 2D cell cultures can therefore be insufficient model systems. In recent years 3D organotypic culture models such as the InSphero 3D Insight™ Microtissues and multicellular tumor spheroids have gained popularity as tools for research, drug discovery and toxicity studies. Several studies have shown the benefit of such models over 2D culture.

Due to their size 3D tissues pose characteristic challenges to any imaging system but benefit greatly from confocal imaging. As the sample is thick compared to the focal depth of the lens, spinning disk confocal systems perform better when the pinhole distance has been increased to minimize pinhole crosstalk, as it is the case for the Opera Phenix™ HCS system.

As 3D tissues are grown in aqueous media and the image quality is greatly improved when using water immersion lenses.

Related Application Notes

Application Note: Imaging Bile Canaliculi in 3D Liver Microtissues using the Opera Phenix High-Content Screening System

Application Note: Quantitative Analysis of 3D Microtissue Growth and Biomarker Intensity using the Operetta High-Content Imaging System

Application Note: Cytotoxicity Studies on 3D Primary Liver Microtissues

Application Note: Quantitative Analysis of Embryoid Bodies Using the Operetta High-Content Imaging System

Application Note: 3D Analysis of Cell Invasion using the Operetta High-Content Imaging System

Publications using PerkinElmer High-Content Screening Products

• Anastasov, N., et. al. (2015). A 3D-microtissue-based phenotypic screening of radiation resistant tumor cells with synchronized chemotherapeutic treatment.BMC Cancer, 15(1), 466. doi:10.1186/s12885-015-1481-9
• Bhatia, M., Lovitt, C. J., Raninga, P. V, Avery, V. M., Di Trapani, G., & Tonissen, K. F. (2016). Expression of the thioredoxin system in an in vivo-like cancer cell environment upon auranofin treatment.European Journal of Cell Biology. doi:10.1016/j.ejcb.2016.08.003
• Ekert, J. E., Johnson, K., Strake, B., Pardinas, J., Jarantow, S., Perkinson, R., & Colter, D. C. (2014). Three-dimensional lung tumor microenvironment modulates therapeutic compound responsiveness in vitro-implication for drug development.PloS One, 9(3), e92248. doi:10.1371/journal.pone.0092248
• Falkenberg, N., et. al. (2015). Additive impact of HER2-/PTK6-RNAi on interactions with HER3 or IGF-1R leads to reduced breast cancer progression in vivo.Molecular Oncology, 9(1), 282–294.
• Falkenberg, N., et. al. (2016). Three-dimensional microtissues essentially contribute to preclinical validations of therapeutic targets in breast cancer.Cancer Medicine, 5(4), 703–10. doi:10.1002/cam4.630
• Garvey, C. M., et. al. (2016). A high-content image-based method for quantitatively studying context-dependent cell population dynamics.Scientific Reports, 6, 29752. doi:10.1038/srep29752
• Letzsch, S., Boettcher, K., Kelm, J. M., & Messner, S. (2015). Quantifying Efflux Activity in 3D Liver Spheroids.Genetic Engineering & Biotechnology News, 35(7), 14–15. doi:10.1089/gen.35.07.08
• Li, L., Zhou, Q., Voss, T. C., Quick, K. L., & LaBarbera, D. V. (2016). High-throughput imaging: Focusing in on drug discovery in 3D.Methods (San Diego, Calif.), 96, 97–102. doi:10.1016/j.ymeth.2015.11.013
• Reid, B. G., et. al. (2014). Live multicellular tumor spheroid models for high-content imaging and screening in cancer drug discovery.Current Chemical Genomics and Translational Medicine, 8(Suppl 1), 27–35. doi:10.2174/2213988501408010027
• Sadok, A., et. al. (2015). Rho kinase inhibitors block melanoma cell migration and inhibit metastasis.Cancer Research, 75(11), 2272–84. doi:10.1158/0008-5472.CAN-14-2156
• Shelper, T. B., Lovitt, C. J., & Avery, V. M. (2016). Assessing Drug Efficacy in a Miniaturized Pancreatic Cancer In Vitro 3D Cell Culture Model.Assay and Drug Development Technologies, 14(7), 367–80. doi:10.1089/adt.2016.737
• Sidarovich, V., Adami, V., Gatto, P., Greco, V., Tebaldi, T., Tonini, G. P., & Quattrone, A. (2015). Translational downregulation of HSP90 expression by iron chelators in neuroblastoma cells.Molecular Pharmacology, 87(3), 513–24. doi:10.1124/mol.114.095729
• Wenzel, C., Otto, S., Prechtl, S., Parczyk, K., & Steigemann, P. (2015). A novel 3D high-content assay identifies compounds that prevent fibroblast invasion into tissue surrogates.Experimental Cell Research, 339(1), 35–43. doi:10.1016/j.yexcr.2015.10.003
• Zhou, Q., et. al. (2016). Topoisomerase IIα mediates TCF-dependent epithelial-mesenchymal transition in colon cancer.Oncogene. doi:10.1038/onc.2016.29