GENERAL INSTRUCTIONS FOR LAB SAFETY AND PROPER FUNCTION OF THE LAB

(Please read carefully)

Rheumatology and Clinical Immunology laboratory

1. CONTAMINATION RISK

  • Always keep your bench CLEAN to avoid contamination. Clean surface of your bench with 70% ethanol.
  • Make your own buffers and reagentsi.e. PBS, culture mediums. DO NOT USE OTHER PEOPLE’S!!!!! You may contaminate their medium or it might be already contaminated.
  • Mediums do not last forever! Always write the date that you prepared the medium and clearly label it with your name and content.

Example

  • When you make mediums/ aliquots it is wise to use parafilm and seal the cap.
  • Always clean hood with 70% ethanol before you work and when you finish!Respect the next person who works after you. Turn on UV light in the hood when you have finished workingwith highly contaminating agents.
  • Stop and THINK!!!!!!! Most of the times contamination is a result of careless actions

Example :

a. When you open a sterile bottle of culture medium outside the hood IT IS NOT STERILE ANY MORE!!!

b. Sterile Tips that are opened outside the hood cannot be used again inside the hood!!!

c. when you are pipetting be careful not to touch the tip to any surfaces.

d. Avoid pipetor contamination DO NOT EXCEED INDICATED VOLUME. If you are using P200 NEVER use it for volumes greater than 200μl.

2. SAFETY ISSUES

  • IF YOU ARE NOT SURE ALWAYS ASK!!!!
  • Always report spills, broken glass etc.
  • Always assume that biological fluids are infectious and use GLOVES
  • When you are disposing needles do not take out the cover! DISPOSE NEEDLE WITH THE COVER!
  • Check labels of reagents that you have not used before and ask your supervisor about handling.

Example : Ethidium bromide can cause mutagenesis. NEVER use it without gloves or inhale it.

  • Discard waste in appropriate containers!
  • CENTRIFUGES: Improperly used or maintained centrifuges can present significant hazards to users. Failed mechanical parts can result in release of flying objects, hazardous chemicals and biohazardous aerosols. The high speed spins generated by centrifuges can create large amounts of aerosol if a spill, leak or tube breakage occurs. To avoid contaminating your centrifuge:
  • Avoid filling tubes to the rim.
  • Ensure that the centrifuge is properly balanced.
  • Do not open the lid during or immediately after operation, attempt to stop a spinning rotor by hand or with an object, or interfere with the interlock safety device.
  • Do not forget the C/F at 4oC!!! If you do not use it anymore RAISE the temperature.
  • At the end of the day leave C/F lid half-open to ensure evaporation.

3. ORDERS / STORAGE

  • ALWAYS NOTIFY IN ADVANCE WHEN REAGENTS/ ANTIBODIES/ PLATES etc ARE ABOUT TO FINISH!!! NOTIFY Dr CHOULAKI WHEN STOCKS REACH 1/3.
  • When you place an order always write on the board and notify Dr Choulaki :

A. the catalogue number

B. the name of the product and

C. company name

  • Always check STOCK places before ordering a new antibody, reagent etc
  • When you borrow reagents from other labs make sure you RETURN them when the new arrival comes.
  • Reagents that are for common use in the lab should be stored in designated places. DO NOT FORGET THEM IN RACKS, BENCHES etc. Other people need them as well!!!!!
  • NEW ARRIVALS should be written on the board otherwise the person who ordered the reagent may not be informed.
  • ALWAYS LABEL BEFORE STORING!!!!!!!!!!This is VERY important especially for long-term storage of samples.

Example:

Tubes labelled as C1, C2, C3, C4 and stored at -80oC

What is in the tube???????????????

  • Always write content ex. RNA
  • Source ex. Human, mouse?
  • Concentration? (optional but make sure you know that it is written in your lab book)
  • Day of extraction
  • Store them in your own labelled box

3. BEFORE LEAVING

  • Perform a safety check! Check that you have turned offgas, water, electricity, vacuum and compression lines, heating apparatus and PCR machinesAND COMPUTERS!!!.
  • Return unused materials, equipment and apparatus to their proper storage locations and to other labs ASAP if you have borrowed equipment.
  • Decontaminate any equipment or work areas that may have been in contact with hazardous materials.
  • Leave behind protective clothing (lab coats, gloves, etc.) when leaving the laboratory
  • Label, package and dispose of all waste material properly
  • DO NOT THROW BIOHAZARD MATERIAL IN THE SINK!!!
  • The dangerous sanitary refuse (RED BOX) is for infectious substancesONLY i.e infectious blood, etc. Do not throw anything in there!!
  • Close and lock the door of the laboratory if you are the last one to leave.