Recent fluorescence microscopy allows for high-throughput acquisition of 5D (X, Y, Z, T, and Color) images in various targets such as cultured cells, 3D spheroid/organoid, and even living tissue with single-cell resolution. The technology is considered promising to augment insights on heterogeneous features of both physiological and pathological cell phenotypes, for instance, distinct responses of cancer cells to anticancer drug treatment. Here we overview microscopic applications to capture live cell events for different types of targets, together with a couple of proof of concepts. The 2D live imaging will be exemplified by a FRET-based time-lapse cultured cell imaging, and 3D tissue imaging protocol will be complemented with a method for mouse skin live imaging.
Keywords: 2D/3D-cultured cell observation; Fluorescent probes; Living mouse observation; Microscopy; Single cell imaging.