S1 - Cell culture guidelines

Getting started:
- Turn on hood, open bench ⇒ needs several minutes to establish flow, wait for green light
- Tie back hair, push back lose sleeves; disinfect hands & arms, put on gloves (can be reused)
- Wipe down bench and window with 70% EtOH
- Gather all material you need, spray with 70% EtOH and wipe down before placing under hood
- Arrange items under hood towards the back of the bench → Keep in mind that you should work a minimum of 15cm inward from the outside edge of the laminar airflow hood; also, leave bottles closed until use and don’t move over open containers; no pouring, use pipettes!
- Always open sterile plastic consumables under the hood
Before you sit down & when you are done: If necessary
- Stock up pipettes and falcons (also glass pasteur pipettes)
- Refill EtOH bottles (also canister, if empty)
- Check vacuum bottles
- Turn vacuum on or off, if applicable
General lab maintenance/ behaviour:
- Minimize waste whenever possible, plan experiments accordingly
- Clean spills immediately with 70% EtOH
- Let appropriate person know if things are running low: e.g. Jan for plastic consumables, Marleen for PFA, Claudia/Susi for Trypsin, FCS/CS etc., Steffi for general S1 issues...
- Thaw new trypsin bottle for next person if current bottle is close to empty
- Fill up glass pasteur pipettes if you emptied a canister ⇒ stick on “heat-dry” tape & put on cart
- Empty trash can if ¾ full, lightly close with tape & place on cart for autoclaving ⇒ use 2 new autoclave bags!
- Collect polystyrene waste (f.ex. pipets w/o filter, cell culture dishes and multiwell plates) in SEPARATE trash can
- Make sure clip on vacuum hose is on completely –> or else pump keeps running!
- Plug in pipette boy if battery is low
- Close bench when you step away for an extended period („stand-by mode“)
- Vent vacuum when you are done (but it‘s not the end of the day yet)
- Turn off computer AND monitor when done using, turn of microscope light when done using
- If fluorescence lamp was used, set timer for 30min ⇒ Should not be switched back on again within 30min!
- Turn on UV light under bench for 30min if a contamination is suspected! (set timer for 30min)
!!! Leave it how you would like to find it !!!