## Materials and Methods
### Device Preparation for Cell Seeding (Static) and \(\upmu\)Fluidic Operation (Flow)
#### 2.2.2 Connecting Culture Wells and Initiating Fluidic Conditions (Flow)  
First, remove the spent media from the cell seeding and acclimation process. Next, add 350 \(\upmu\)L of fresh media to the sink well (well 12), 100 \(\upmu\)L of fresh media into columns 2-11, filling the wells from right to left (Column 11, 10, 9... 2), and 400 \(\upmu\)L of fresh media into the source well. Return the filled platform to a humidified 37 \({}^{\circ}\)C incubator, flow begins  
\begin{table}
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**Description** & 96-well geometric footprint (1/2 area). 8 replicate flow channels (A-H) with linked wells. \\  & External Dimensions: & Lab equipment agnostic. Compatible with most plate readers and most imagers. \\  & Disposable Unit: & 1\(\times\) use. Amendable application models such as modular cell-system configurations. \\
**Cell Alignment \& Analysis** & Biologic support and functions. \\  & Accessibility: & Cell harvesting (cellular analysis.). Media aspiration (fluid analysis). \\  & Cell Phenotype: & Single cell phenotype. Multi-cell phenotypes. Cell-cell signaling. \\  & Fluorescence Detection: & Optically clear wells. Image through plate bottom. Plate shall not auto-fluoresce. \\
**Format** & (x, y) well locations mimic traditional positioning. Z-height varies across 4 mm vertical. \\  & Growth Area per Well: & Surface area is 0.167 cm\({}^{2}\). 2D, flat surfaces. Side walls have edges for manufacturing. \\  & Each Channel Includes: & Dose well (1); fluid regulator well (1); culture wells (9); waste well (1). \\  & Device Composition: & Dosing wells (8); fluid regulator wells (8); culture wells (72); waste wells (8). \\  & Z-Height Offset: & Wells 2–11, surfaces have 0.5 mm z-height offset amid adjacent (serially linked) wells. \\
**Channels for Fluid Flow** & Paths connect wells. Capillarity involves both 3-wall and 4-wall designs; air venting. \\  & Fluid Traverse: & Passageway transport initiates in well 1 and terminates at well 12. Unidirectional flow. \\  & Dose Well (Well 1): & Max vol. 500 \(\upmu\)L. Fluid flow passageway from bottom of well 1 into top of well 2. \\  & Regulator Well (Well 2): & Max vol. 100 \(\upmu\)L. Force equilibrator w/ fluid hydrodynamic. Fulcurum amid flow paths. \\  & Culture Wells (Wells 3–11): & Working vol. of 30–100 \(\upmu\)L; 1/2 area of 96-well format; 4 mm well height. \\  & Sink Well (Well 12): & Max vol. 350 \(\upmu\)L. Evaporation wick (syphon) pulls fluid from well 1 to well 12 (1-way). \\  & Device Fluid Volume: & Per channel working volume is 1850 \(\upmu\)L (1.85 mL). Per device (8 channels), 14.8 mL. \\  & Fluidic Mixing in Wells: & Rapid mixing. Involves surface tension, laminar flow, well shapes, and fluid dispersion. \\  & Flow Forces: & Gravity driven, hydrodynamics, capillarity, wicking. \\
**Materials in Fabrication** & Polystyrene, optically clear wells, conventional surface tension, base system with lid. \\  & Thickness of Platform: & 0.5 mm polymer thickness; parameter for injection molding and QA/QC non-warping. \\  & Smoothness: & Well bottoms. Avoid bubbles in resin (injection mold flow rate). Contamination free. \\
**Manufacture Units** & Mass produced, 2 components, injection molding. Large quantity production capacity. \\  & Manufacture Components: & Lid (1 per unit). Row cover (8 per unit). Wick (8 per unit).  
Assembly of Units: & Ultrasound welding of molded parts. No glue. No adhesive. No bonding contaminants. \\  & Surface Treatment: & Tissue culture plastic (TCP). Can be surface coated (e.g., proteins, non-binding, etc.). \\ \hline \hline \end{tabular}
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Table 1: \(\upmu\)Fluidics Culture Device—Attributes and Infrastructure.  
automatically. The same process is followed regardless of whether it is a cell line, primary cells, or different cell phenotypes seeded in separate multiwell chambers within the device.