Schematic of cell seeding and immunostaining procedures. (A to H) shows the configuration of device, tubes, and plugs for seeding the device with cells: (A) priming the device with a solution of extracellular matrix (ECM) proteins, (B) coating with ECM proteins, (C) exchanging ECM and cell medium tubes, (D) flushing excess ECM proteins out of the device, (E) inserting the pipette tip containing cells, (F) flowing cells into the device, (G) exchanging the pipette tip with a plug, and (H) incubating the device while cells attach and spread. (I to L) shows the basic steps involved in in situ immunocytochemistry: (I) using negative hydrostatic pressure to introduce an immunocytochemical reagent into the device, (J) incubating the reagent for the desired period of time, (K) generating a liquid bubble, and (L) washing out the reagent with PBS. The inlet and outlet sides of the device are indicated with I and O, respectively, and the arrow indicates the direction of flow, if any.