Safe imaging of live samples

Live sample imaging is a powerful technique and is commonly performed on many of the LMCF systems. The guidelines presented below will ensure you and all other users of the facility are safe when such experiments are carried out.

Overview

There are different BioSafety Levels (BSL) for live samples used in research

  • BSL1 - Yeast, E coli etc not known to cause disease in humans
  • BSL2 - All human cells (including cultured cell lines like HeLa) which may contain pathogens (eg HIV or HBV)
  • BSL3&4 - Very dangerous e.g. Ebola, airborne pathogens. You CAN NOT image such samples in LMCF - you need to dress like an astronaut and do this somewhere specially designed for the purpose.

Standard operating procedure for BSL2 samples

1. Samples must be prepared in your lab using the appropriate TC hoods and safety procedures

2. The culture dishes must be sealed with parafilm to minimize the risk of spills on the microscopes and the outside of the dish wiped with an appropriate disinfectant that is effective against the agent

3. Samples must be carried to the LMCF room in a closed secondary container with absorbent materials in the bottom of the container

4. Use gloves when moving the dish onto the stage, remove gloves before touching the microscope or computer.

5. After imaging, the samples must be removed from the microscope and the microscope stage should be wiped down with tissue soaked in 70% ethanol (or another disinfectant that is effective against the agent). Any spills must be cleaned thoroughly immediately with an appropriate disinfectant that is effective against the agent . Please inform LMCF staff if you do spill something.

6. All samples must be taken back to the host lab for disposal, transported as 3 above. LMCF is not able to handle biohazardous waste for you.

7. Wash hands with soap on finishing the above

BSL2 actually encompasses a surprisingly broad range of samples (e.g. HeLa cells to live HIV!) so please consider if there are other things necessary for your samples.

Questions

If you aren't sure what to do please ask. For any procedure differing from the workflow above we can receive guidance from the Duke Safety Office.