High-Content Image Analysis

High-content imaging (HCI) and image processing of cells grown in 3D pose a significant challenge because 3D cells are not grown in a single focal plane, and the cell culture systems are often incompatible with traditional microscopes. Confocal microscopy is an important tool for imaging 3D cells due to its ability to acquire high definition images at various optical sections. However, the low scanning rate and depth induce low throughput in image acquisition and also incur additional problems such as photobleaching or phototoxicity [1–3]. To alleviate these issues, miniaturized 3D cell cultures on a micropillar/microwell chip platform have been demonstrated, which facilitate high-throughput spheroid cultures in hydrogels while offering better imaging capabilities. The whole sample depth can fit within the focus depth of a normal objective lens due to the small dimensions.