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In combination with imaging and molecular\/biochemical techniques, 3D organoid models have contributed substantially to our knowledge about the cellular and molecular mechanisms that regulate the growth of tumors and invasion into the surrounding tissue. We here outline a set of protocols that describe culturing of cancer-derived organoids in 3D matrices and various strategies that allow modeling of tumor growth, tumor cell penetration into basement membranes, and invasion into Collagen I-rich ECM. Furthermore, we specify protocols for subsequent handling of organoids cultured\u00a0in 3D ECM for confocal microscopy and analysis of gene expression at the protein and mRNA level. Although we here use breast cancer-derived organoids, these protocols can be directly applied or adapted for organoids derived from other cancer types or healthy tissues. Thus, in addition to investigating cell behavior of multiple cancer types, the combination of protocols described here may be used to study processes such as cell differentiation and migration during homeostasis and normal development.<\/jats:p>","DOI":"10.1007\/978-1-0716-2887-4_17","type":"book-chapter","created":{"date-parts":[[2023,1,18]],"date-time":"2023-01-18T17:03:06Z","timestamp":1674061386000},"page":"281-303","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Multimodal Techniques to Study Tumor Growth, Basement Membrane Breaching, and Invasion in 3D Matrices"],"prefix":"10.1007","author":[{"given":"Daan","family":"Smits","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antoine A.","family":"Khalil","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2023,1,19]]},"reference":[{"issue":"12","key":"17_CR1","doi-asserted-by":"publisher","first-page":"a005066","DOI":"10.1101\/cshperspect.a005066","volume":"2","author":"MA Schwartz","year":"2010","unstructured":"Schwartz MA (2010) Integrins and extracellular matrix in mechanotransduction. 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