Hosuk:Cell fixation in culture dish
Jump to navigation
Jump to search
- For cells in MatTek glass bottom dish(P/N : P35G-1.5-14-C)
- NO aspirate media. add 200 ul formalin(~39% formaldehyde) to 2 ml media directly,
- incubate for 15 min at 37C
- aspirate
- quickly add pre warmed 10% formalin in PBS
- incubate at 37C for 15 min
- aspirate
- wash with PBS 3 times at RT
- incubate in 70% etoh for 1 min or store in 70% etoh up to a week at 4C
Jonathan's protocol[edit]
- 100% confluence, and could wait one more day, and split
- When cells are transferred from 6 well plate
- 500uL of Trypsin, incubate 5min
- Add 500uL media, and mix well, and
- 100uL media in 96 well, or 300~350uL media in glass bottom dish
- Add 60uL of cell in 96 well or 200uL in glass bottom dish
- Culture one or two more day
- Fixed cells just using 10% Formamide
- Aspirate media
- Add 50uL of 10% Formamide (200uL for glass bottom)
- Incubate for 15min
- Wash with PBS three times
- Store in Ethanol
PGP1F cell culture in 96 well plate[edit]
- Add 1mL Trypsin in T25 flask (about 90~100% covered)
- Add 4mL media (total 5mL)
- Add 100uL cell (from 5mL in T25 flask) to each well in 96 well plate (total 16 wells)
- Add 200uL media to each well, and gently mix