Table 1.
Neuron and Astrocyte Experimental Task List by Day
| Time Relative to Neuronal Prep | Neurons | Astrocytes |
|---|---|---|
| 1.5 – 2 weeks prior | Culture Primary Astrocytes | |
| 2 days prior | Prepare Coverslips | |
| 1 day prior | Wash & Coat Coverslips with Substrate |
|
| Day 0 | Prepare & Plate Primary Neurons | |
| Day 1 | Change neuronal medium | |
| Day 2 | Treat with ARAC | |
| Day 3 | ||
| Day 4 | ||
| Day 5 | Change 1/3 neuronal medium | |
| Day 6 | Pass experimental astrocytes |
|
| Day 7 | Change medium | |
| Day 8 | ||
| Day 9 | Change 1/3 neuronal medium | Change medium |
| Day 10 | Serum Deprive Astrocytes | |
| Day 11 | Treat Astrocytes | |
| Day 12 | Stop Treatment | |
| Co-culture neurons and astrocytes | ||
| Day 13 | Stop co-culture | |
| Fix neurons & store or begin Immunolabeling |
||
| After Day 13 | Complete ICC labeling Mount Image, deconvolve, analyze |
|