Figure 2

A) Open cryostage, the lid (1) is detached, the i3C (2, cloudy structure) is attached via its adaptor (3, metal frame around the i3C) to the cooling element (4, circular silver body). (B): The lid is closed. Dry nitrogen gas pipe is in place (5). The sample in the i3C is ready to undergo a freezing - thawing cycle. Centers of the lid's window (through which images are acquired - light coming from the condenser is seen) and the i3C's bottom are overlapping and situated on the microscope optical axis. (C) Cross section of the i3C placed on the cryo-stage silver cooling element (1) ready for experiment. The i3C polycarbonate body (2) hosts the glass picowells chip (3) which is secured to its place by bonding. Bottoms are aligned for a perfect contact with the flat cryo-stage. Cells and media (4) which were previously loaded into the i3C are trapped in the cell chamber and the flow conduit due to the capillary action of the sliding cover slip (5). The cryo-stage lid is then closed so that the center of its window (6) and microscope objective (7) optical axis are overlapping to facilitate microscopic observation of cells in the i3C. Note that for the sake of clarity the colors of the i3C components here match those appearing in Figure 1A.