9. Aspirator jars must be emptied before the pumps get flooded. Put some hycolin into the jars before reconnecting.
10. Waste must be disposed of correctly:
• wrappings, paper towels and gloves into the yellow bin bags only.
• anything that has been in contact with cells, media or serum must be disposed of in the autoclave waste. Seal liquids in tubes/flasks or rinse down the sink. It is your responsibility to empty full autoclave bags. The waste should be double bagged, tied at the corners so that steam can penetrate; complete and attach an autoclave waste sheet. Tied bags should
be placed in the containers outside established cell culture for the porters to take away.
• sharps bins should only contain pipettes, needles, tips and broken glass, fix and lock the lid and leave by the door in the link corridor for the cleaners to take away.
• Never endanger the support staff by disposing of waste inappropriately.
11. Users are to supply their own sterile tips and pasteur pipettes.
12. When you have finished culturing, remove everything from the hood, spray the hood with dilute Virkon, wipe down and then spray with 70% IMS.
13. If you are the last person to use the facility, shut down the hood, ensure its empty (remove disposal pots) insert the night-door and switch on the UV light (where applicable) (1 hour is sufficient time for the UV light.). Make sure the waterbath is switched off.
14. Treat culture medium with hycolin prior to pouring down the sink; & rinse down with water.
15. Centrifuges are a potential source of contamination, in case of spills, clean thoroughly with an appropriate disinfectant.
16. Only items used regularly for culture should be stored within the cell culture facilities. "Junk" makes it difficult to keep things clean and is a safety hazard- store it elsewhere!
17. The hycolin solution in the pipette stacks must be changed every Monday morning. The old solution must be poured down the sink through a sieve and the tips and glass in the sieve must then be disposed of in the sharps bin. NB if pipettes are used for adenoviral work,they should be soaked in virkon prior to transfer to hycolin.
18. If you discover an infection in your cultures you must immediately remove the cultures from the facility, decontaminate as appropriate, dispose of them in an autoclave bag and inform other users of the problem. Track down the source of the infection, decontaminate and dispose of that too.
19. To facilitate communication regarding specific problems or if an item needs restocking, there are marker boards in cell culture facilities which Caroline will check 2/3 times a week.It is the responsibility of the cell culture users to maintain their own supplies of trypsin, lglutamine, and pen-strep etc.
D. Hood Booking and Designation (Communal facilities only)
For communal facilities adjacent to Labs 1 and 8, there is a diary for Hood booking for each hood. No-one may book more than three weeks in advance. Priority always lies with the person booked in - -and this person is always responsible for the condition in which the hood is left.
Primary culture adjacent lab 1 has a double hood for human blood work and a single hood for rodent primary culture. When not in use for rodent culture, the single hood may be used as an overflow for human blood work but animal users have booking priority.
The double hood for human blood work may be used as an overflow for rodent work, BUT those carrying out human blood work have booking priority and the hood must be cleaned thoroughly after use.
Established culture (adjacent lab 8) is maintained as a mycoplasma-free facility (see below) with registered projects for adenovirus work.
NB Embryonic stem cell work will occasionally be carried out in the smaller of the two rooms.When ES cell work is planned 4 weeks notice will be given and all other cells will be moved to the larger room. The smaller room will then be shut down, cleaned and formaldehyde 'bombed'. Once this has occured then NO OTHER cells may be cultured in that room whilst it is in use for ES cell work.
