SEM 관찰용 세포 고정방법 (Cell Fixation)

Cell Seeding & Cell Attachment Assay

1. Fix the scaffolds into desired well plate in clean bench

2. Sterilize the scaffolds by ethanol for 1-2 hours if applicable

3. Keep the scaffolds under UV light for sterilization for around 90mins

4. Wash the scaffolds 3x with PBS for 2min

5. Incubate the scaffolds with complete medium (medium + serum+ antibiotics) for 1-2 hours at 37℃

6. Aspirate medium and seed scaffolds with the desired amount of cells. Incubate for 15-30 mins to       allow for attachment

7. Add appropriate amount of medium and incubate for desired amounts of time (e.g., 1, 3, and 7days)       while changing medium every 48 hours

8. Aspirate medium and wash cells 3x with PBS.

9. Add 300-500ul of 3-5% glutaraldehyde solution and keep at 4℃ for 1-2 hours

10. dehydrate cells with a series of graded 25, 50, 75 and 95% ethanol, each for 5-10 mins

11. Dry the plates for 1~hour in a 37℃ oven or overnight at room temperature