This protocol demonstrates how to create functionalized coverslips for inverted microscopy. These coverslips can be used for cell culture or Drosophila whole mounts. Pay attention to the coverslip thickness and that it matches your objective/settings.
Always label batches with coverslip thickness.
This protocol assumes the etching and coating is done in Coplin jars.
| Item | Item article number | Distributor | Vendor |
|---|---|---|---|
| APTES | 440140 | Merck | SigmaAldrich |
| UltraPure™ DNase/RNase-Free Distilled Water | 10977035 | ThermoFisher | Invitrogen |
| Glacial acetic acid | A6283-100ML | Merck | SigmaAldrich |
| Cover glass Borosilicate | 631-0124 | VWR | VWR |
Clean coverslips in 1M KOH in for 20min.
(make 1 L of 1M KOH by dissolving 56.11 g of KOH in 1 L of deionized water).
Rinse three times in nuclease-free water.
Rinse one time in 100% methanol.
Let the coverslip dry from the methanol and then immerse in APTES, (3-aminopropyl)triethoxysilane, for 2 min:
| Reagent | Amount | Volume | Final concentration |
|---|---|---|---|
| 100% Methanol | 55.2 mL | ||
| Acetic acid | 3 mL | 5% vol/vol | |
| APTES | 1800 µl | 3% vol/vol | |
| TOTAL: | 60 mL |
Rinse three times in nuclease-free water.
Dry the coverslips in a dark and desiccated environment in room temperature.
Store dry or use immediately.