Figure 2

Flyspresso components and design. (A) Photograph of the Flyspresso robot (_24W.iam), composed of (from top to bottom) the System Piston, Aspirate Manifold, Dispense Manifold, Separation Manifold, Microplate, and the Heating and Shaking Device. (B) The syringes and the chemical manifold are controlled by an Arduino Mega 2560 microcontroller, which communicates internally with a python interpreter on a computer with the specific protocol. (C) The Chemical Manifold (24 W CHEMICAL MANIFOLD ASSY.iam) can be expanded to the number of channels needed for each experiment. Each chemical channel is controlled by a solenoid valve, which is dictated by the Arduino microcontroller. Chemicals not in use are held via vacuum, and the desired chemical is pulled into Flyspresso when the solenoid valve switches to ambient air (see also Fig. 3). (D–H) Exploded-view of Flyspresso. (D) Schematics of the System Piston. Color schematic from red to violet corresponds with the piston’s position when pulling liquid into the system (24 W SYRINGE SYS.iam). The System Piston moves a System Liquid to control the other syringes. Movement is controlled by a gas and vacuum supply (see also Fig. 3). (E) (Left) The Aspirate Manifold (24 W ASPIRATE MANIFOLD.ipt) sits between the Chemical Syringe Pumps. (Right) Top view of the Aspirate Manifold laying on the Dispense Manifold. Chemical waste is pulled through the Aspirate Manifold to a waste container. (F) (Left) The Dispense Manifold (24 W DISPENSE MANIFOLD ASSY.iam) holds a Priming Syringe Pump (P) and 6 Chemical Syringe Pumps (i–vi). (Right) Top view of the Dispense Manifold shows the chemical pathway to either the Priming Syringe Pump (P) or the 6 Chemical Syringe Pumps (i–vi). (G) (Left) The Separation Manifold (24W SEPARATOR ASSY.iam). (Right) Top view of the Separation Manifold. The Separation Manifold is equipped with 24 separator tips, which aspirate liquid from the upper interface to Separation Waste. (H) The custom microplate (24 W MICROPLATE ASSY.iam) sits below the Separation Manifold and holds 6 Transplates (yellow) (24W TRANSPLATE.ipt) for a total of 24 samples. Seplate Attachments (gray) (24W SEPLATE.ipt) sit above the Transplates and enclose the embryos, separating the upper and lower liquid interfaces.