A key feature of organoid culture involves embedding cells within an extracellular matrix that permits the cells to grow in three dimensions into large, complex structures with varying morphologies. However, these features can also make quantification of culture health and proliferation challenging. Unlike 2D monolayer cell cultures, organoids do not proliferate as single cells, which can make cell counting and viability quantification approaches difficult. Here, approaches including commercially available kits to quantify metabolism or ATP and the trypan blue dye exclusion assay were utilized. The results were compared with label-free imaging approaches from multiple instrument platforms. Additionally, a small-scale toxicity assay was performed with various chemotherapy drugs to assess the assays. Results varied between models, donors, tissues, and cancer types. Overall, to accurately assess the growth of such complex organoid cultures, significant optimization and validation may be required. Depending on the specific application, either imaging based or cell-based assay approaches may be suitable.