Inverted microscopy is a very popular technique for live cell imaging. Here, living cells are observed through the bottom of a cell culture vessel. This technique has several advantages over upright microscopy. Most cells naturally sink to the bottom of the vessel and—if no suspension cells are used—adhere to the surface, meaning that they are spread across one focal plane. When growing in vessels that are suitable for inverted microscopy, cells have access to larger amounts of medium than if they were squeezed between a coverslip and a slide. In addition, sample access from the top is possible, for medium exchange or micropipettes, for example. Another very important advantage is sterility: as there is no contact between the objective and the sample, sterile working conditions are guaranteed.