Image cytometry uses microscopy and image processing to assess cell properties in biomedical research, drug discovery, and clinical diagnostics. In many workflows, it is desirable to isolate individual cells or cell clusters into separated chambers to observe their behaviour free from the interference of other cells. These small chambers, referred to as nanowells, typically have volumes in the nano-liter range and lateral dimensions from 25-500 µm. One drawback amongst current nanowell products is that they are generally fabricated out of a single-material which presents a problem when providing a growth surface for adhesion cells and usually introduces optical limitations. Here, we are introducing ImageCyte’s innovative glass-bottom nanowell imaging plates designed to meet these demands. Incorporated into standard 96 and 384 well formats, each well features a grid of nanowells fabricated directly onto the coverslip-thin glass bottom. The nanowells come in various sizes and configurations, catering to the culturing requirements of suspension and adhesion cells; as well as an optional low-attachment coating for working with spheroids.