Live cell microscopy
Preparation of culture dishes
- In 3.5 cm culture dishes (Sarstedt 83.1800 35x10 mm) a hole is drilled with 14 mm in diameter (will be done by the uni garage)
- Before sticking the coverslips (Menzel 24 mm, Stärke 1,5; CB00240RAC20MNZ#0) to the dishes spray them with 70% ethanol and wipe with a KimWipe tissue
- The coverslips are pretreated as following:
- Wash with a mixture of 50% v/v Aceton and 50% v/v EtOH
- Wash with 1xPBS
- Wash with MilliQ water
- Dry and sterilize at 120°C over night
- Stick the coverslip with Fixogum (Marabu) to the culture dishes
- Dry for two days in the dark
Coating of culture dishes and seeding of cells
- Culture dishes are sterilized for 5 min at 850 W in a microwave (add a bottle of dest. water with a loosely tightened lid as the microwave must not be used dry)
- Culture dishes are coated depending on the cell type (1-2 h at 37°C or at 4°C over night)
- 1∙105 cells per dish were seeded (also depending on the cell type) in 2 mL of culture medium
- Before starting imaging replace culture medium to colourless medium (wash with colourless medium first)
Preparation of the microscope
- Preheat the incubation chamber (2-3 h before starting the experiment)
- Connect CO2-supply