ESO wikiadmin: Created page with "=== 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 Dr..."

2025-05-20T08:39:41Z

Created page with "=== 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 ** Dr..."

New page

=== 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∙10<sup>5</sup> 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 CO<sub>2</sub>-supply

Live cell microscopy - Revision history