Reagents and cell
This protocol is extracted from research article:
Autonomous microfluidic actuators for periodic sequential flow generation
Sci Adv, Apr 19, 2019; DOI: 10.1126/sciadv.aat3080

For the experiments represented in Figs. 3 and 4, we used a 2 mM blue (quinine hemisulfate salt monohydrate, Sigma-Aldrich) fluorescent dye in 5% sulfuric acid, 400 μM green (fluorescein sodium salt, Sigma-Aldrich) fluorescent dye in deionized water, and 400 μM red (rhodamine B, Sigma-Aldrich) fluorescent dye in 5% ethanol. For the experiment shown in Fig. 5, human umbilical vein endothelial cells (HUVECs) were cultured in T-flasks using standard cell culture methods. The cell seeding channel was coated with 0.4% (v/v) fibronectin in phosphate-buffered saline (PBS) at 37°C for 30 min. Then, the cell seeding channel was rinsed with PBS, and cell growth medium [Lonza, EGM-2 BulletKit (CC-3156 and CC4176)] was filled. Next, HUVECs were removed from the T-flask and injected into the cell seeding channel. For the attachment of the cells on the channel surface, the device was placed in an incubator with 5% CO2 at 37°C for 2 hours. To stain cell nuclei (Fig. 5), we used a 1 μM fluorescent dye (SYTO 85, Invitrogen) and tris-EDTA (TE) buffer.

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