| Step # | Process Step | Active Time | Wait Time | |
|---|---|---|---|---|
| Day 1 | 37 | PDMS molding | 30 min | overnight |
| Day 2 | 38-45 | PDMS device assembly | 1 hour | 15 min |
| 46-47* | Surface treatment of devices | 15 min | 4 hours - overnight | |
| 48-50 | 30 min | overnight | ||
| Day 3 | 51-60 | Preparing collagen gel | 1 hour | 1 hour during, then 4 hours - overnight |
| 61-66 | Removing needles & rinsing gels | 30 min | overnight | |
| Day 4 | 67-69 | Cell preparation | 10 min | |
| 70-76 | Channel seeding | |||
| 77a, i | Gravity-driven oscillatory flow | 5 min | Application dependent, 14-72 hours | |
| 77b, i-v | Syringe-pump driven unidirectional flow | 30 min | overnight | |
| Day 5** | 77a, ii | Gravity-driven oscillatory flow | 10 min | |
| 77b, vi - xv | Syringe-pump driven unidirectional flow | 1.5 hours | overnight | |
| Day 6** | 78-82 | Permeability assay | 5 min per device | |
| 83-91 | Visualization of cellular junctions and cytoskeleton via immunofluorescence staining | 1 hour | 1 hour - overnight | |
| Day 7 | 92 | 15 min | overnight | |
| Day 8 | 93 | 30 min | 30 min - overnight | |
| 94 | 15 min | 45 min - overnight | ||
| Day 9 | 95-96 | 30 min |
*
Do not proceed to this step unless you plan to continue through step 65 in accordance with the timeline
**
On these days, choose one of the two paths, depending on the experimental design and goals