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Guidelines for using the Cell Culture Room

General Rules for Use of Facilities

1. Everything brought to and stored in this room (in hoods, on/in shelves/drawers, in refrigerators & freezer) must be labeled with your names and your PIs initials. Labeled items should not be moved by anyone else without permission under any circumstances. If items need to be moved or attended to in any other way, contact the relevant lab/person as indicated on the label or the PI of the lab.
2. Refrigerators can only be used to store those items to be used explicitly for cell culture.
3. Individual labs have access to the –80o freezer for the storage of unstable materials. If a machine happens to break, a cylinder needs to be replaced etc, the appropriate steps must be taken immediately to facilitate the repair or replacement of the said items. This does not mean that your lab will be required to cover these expenses. Timely repair avoids inconvenience to others.

Rules for use of Hoods
So you’ve signed-up and you’re now ready to do some work in the hood:

1. Turn on blower and inspect the hood. The hood should have been cleaned by the previous user to avoid contamination problems.
2. Wipe down the surfaces in the hood with an ethanol soaked Kim wipe. Don’t spray ethanol directly into the hood!
3. Check to make sure the sharps container is not filled past the fill line.. If it is, tape, label and replace.
4. When you finish your work, remove any items that you may have brought into the hood (media, pipetter, disposable pipettes, etc).
5. Wipe with ethanol again, turn on the UV light, close hood and turn off the blower.

Rules for use of incubators

1. Everything in the incubators must be labeled with your Name, PIs initials, Date, and Cell Line.
2. Verify the CO2 level in the cylinder attached to your incubator is adequate else report to lab on duty. Do not change the CO2 setting with out consulting other using the incubator.
3. If any problems are identified in using the incubators, report to individual researcher or PI.

Sign-Up Procedure for Hood Use

To facilitate better coordinator between the users of the hoods in the Cell Culture Room, online sign-up sheets have been provided through the www-sched program.

This information will help us to share the room more effectively. Please do not neglect to sign up every time.

Lab Maintenance Duties

Each lab using the room will have the responsibility for maintaining the room/equipment on a weekly basis

The duties are as follows:

Sharps containers: these containers have explicitly labeled “fill lines”. They should not to be filled above these lines. The user or personnel from the lab on duty (“whoever is first on the scene”), must tape the container closed and label with “Biohazard Sharps”, their name, date, and then place the container in the larger biohazard dry waste receptacle in the Cell Culture room for pick-up.

Disposable pipettes: Two soaking containers are provided, to be used alternatively. Pipettes must be completely submerged in 50:50 bleach/water solution until the container reaches capacity. It must then be removed and stored overnight in the “on-duty lab”, the pipettes then dried and put in a box, taped closed and disposed of (regular garbage bin is fine).

CO2 Cylinders: These must be checked periodically throughout the week to ensure that there is enough CO2 in reserve. When the cylinder pressure gauge (not exhaust pressure) read less than 250psi the CO2 should be replaced and a new cylinder ordered through Praxair. When setting up, the CO2 exhaust gage (the feed into the incubator) should never read above 10psi.

Liquid waste collection jars: Jars are located in each hood. They must be emptied when they are no longer clear in the sink, rinsed with tap water, and filled with approximately 300ml of bleach.

Water baths: Their water levels must be checked periodically each week; top up as necessary to indicated level. Anti fungal/algae additive should always be used in the water bath.
Other supplies: Check that there are adequate supplies of sharps containers, bleach, ethanol and biohazard bags.

Training
New users to the Cell Culture Facility should check in with their PI before using this equipment. Graduate and undergraduate students will be required to shadow an approved user from their respective lab before being approved for solo use of this facility.

Key Access:
Access key codes may be assigned by Linda Schmidt to Graduate Students, Post Docs and Staff members only. Have the appropriate form filled out and signed by your PI to gain access to this room.