Guidelines cell culture laboratory

Laboratory protocols

GUIDELINES FOR USE OF CELL CULTURE LABORATORY GADE LABORATORY FOR PATHOLOGY

NB! Anything you wonder about you should ask Jorunn or Dana. Please do not place things at cell culture lab without notifying Jorunn or Dana.

Every person working in the cell-lab should be familiar with the following procedures:

• Dress code in the cell culture lab
  • Use shoes dedicated for use exclusively in the cell culture lab or plastic single-use cover that you find at the entrance of cell culture lab
  • Use blue coats for use exclusively in the cell culture lab
  • Use mask & gloves
  • Know where to find new coats
  • Know where to discharge used coats

NB! There is only one dedicated person that brings to the lab single use plastic cover, masks and gloves – please contact him/her for renewing the stocks in the lab. At the moment is HARSH DONGRE.

• General rules on how to work in the cell culture lab
  • always book your time in advance for the respective hood (schedule on the door)
  • no more than 4 people at the time are allowed in the cell culture rooms
  • keep the doors closed
  • wash your hands before and after starting work in cell culture lab
  • respect the weekly/monthly duty schedule for cleaning the lab

• Know how to use / wash the hoods & incubators.

NB! Always write in the logbook if you use the hoods.

• Know how to check CO2 level and change the flow from empty to full CO2 flasks. Notify Jorunn or Dana if you find empty flasks.

If the incubators are bipping because of low CO2 you have to change the flow of CO2 from the empty to full CO2 flasks. The CO2 flasks are in the storage/ventilation room. The key to that room are on the side of the hoods in the main wet lab. Notify Jorunn / Dana if you changed the flow to the new ones so we order new reserve flasks.

• Know how to check the nitrogen tanks and notify Bendict, Dana or Jorunn if the tanks are bipping.

NB! At the moment Benedict will order nitrogen.

• Know how to use / wash the water bath.

           Wash the water bath once a week with distilled water and add AquaClean.

• Know how to use the microscopes.

Remember to turn off the microscope after use and cover with the plastic hood. Clean up if you spill cell suspension etc. Clean the lenses with lens paper. For the fluorescence microscope:

• • always write in the log book if you use it
  • if you start the lamp, let it in use for at least 15 min, then turn it off
  • check the log book before starting the lamp. Do not start the lamp if the microscope was use less than 30 min before.
• Know how to use the autoclave.

On the autoclave you will find a manual for operating the instrument. NB! Ask Saroj if you are not sure how to use it.

• Know where to get Millipore-filtered water
  • washing room at Molecular pat lab, 2 floor.

• Know how to use the washing machine.

On the washing machine you will find a short instruction. NB! Ask Dana or May Britt if you are not sure how to use it.

• Know how to throw the waste.

Pipettes and flasks should be separated and placed in the cardboard boxes with a red plastic bag inside and marked with an orange sticker: ‘Patologisk’ with the date and name of the dept.: ‘Patologi’. Not more than 2/3 full. Bring them to the container room (just outside the main glass door) after folding up the box. Everyone has the duty to remove any 2/3 full boxes you find in the cell lab. NB! Don’t place any unmarked boxes in the container room! Used medium. All used medium should be sucked in the special containers using the pumps connected with the hoods. When the special containers are full they should be placed in cardboard boxes with a red plastic bag inside and marked with an orange sticker: ‘Patologisk’ with the date and name of the dept.: ‘Patologi’. Ordinary waste (not contaminated) is to be put in the ordinary waste bins (with white plastic bag) in the main lab or in the big waste bin (with black plastic bag) in the corridor outside the cold room. The hospital cleaner will take care of these. Plastic waste: use the cardboard boxes with transparent white plastic bag in the room with the ice machine. Plastic waste should be sorted in in the two boxes that are there under the centrifuge in soft plastic and hard plastic. The hospital will take care of these boxes. Do not throw used gloves in the boxes containing plastic. Contaminated gloves should be thrown in the boxes with red bags inside marked “patologisk avfall”. Used gloves with no contamination are thrown in the ordinary waste bins. Sharp objects (pipette tips, scalpels, needles) should be thrown in small yellow boxes. Place them in a cardbox with a red bag. Mark the boxes with yellow sticker “stikkende/skjærende” and place them in the container room. NB! Don’t place any unmarked boxes in the container room! Cardboard and isopor boxes /Packages you receive: When you empty cardboard and isopor boxes, place them in the container room, not by the paper collection boxes or on the floor. We keep only few isopor boxes in the shelf outside the cell culture lab for use when work on ice is needed. Fillings (isopor) are thrown in a designated waste bin outside the cell lab. Discharge of solvents. Used alcohol should be kept in the fire cabinet in the main lab until the plastic can is full. Then bring the bottle to the histolab. Pour used xylene/acetone in a plastic can and keep this can in the hood (in the mail lab) until it is full. Then throw the xylene in the sink especially put up for xylene waste. This is in the histolab were the ethanol pump is placed. Discharge of broken glass. Notify Jorunn or Dana if you have some broken glass. It has to be thrown in a designated waste bin up in the pathology lab.

• Know how to use the cooling room.

Remember to close the door firmly and switch off the light!

• Know how to store your materials / disposables

Every research group have several designated cabinets in the lab.

• Know where to weight chemicals
  • washing room at Molecular pat lab, 2 floor.

• Know that all chemicals have to be registered on EcoOnline. Register for chemicals (All users should have a read function).

When you order and receive new chemicals: give May Britt information about name and number of chemical, amount, company and how you will use it (open vessel, closed vessel, on bench or in hood,- this is for the risk evaluation).

• Know when it is your turn for weekly duties. Remember to check the lab at least twice a week when it is your turn
  • wash lab instruments & refill the boxes with gloves, masks, shoe covers, etc.
  • discharge the boxes filled with used pipettes, or used cell culture dishes.
  • discharge the yellow plastic boxes filled with sharp objects
  • check the water level in the incubator & wash the water bath
  • check the level of CO2 and liquid nitrogen.

• Participate at the monthly/half year duties
  • wash the incubators & the hoods.
• Participate at the monthly lab meetings.

If you have problems to see the form below, you can also find the self-declaration form here:

• http://link.uib.no/labproto

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