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BL2-level safety at the microscopy core. Users who need to image live human and non-human primate samples MUST read these instructions.
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BL2 users must read these safety policies and take a short training session in BL2 imaging practices with core personnel.
Make sure to read the
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[link] before continuing.
Key Points for BL2 Usage
- Determine whether your sample is BL1 or BL2 (see below). Ask us if unsure!
- BL2 sign and extra preparation are required while imaging BL2 samples
- Don't pipette or add to your samples on the microscope stage!
- Use our local BL2 manipulation area for that
- Transport samples in airtight secondary containers with latching lids.
- MIT users must amend their BRR to include the microscopy core.
- You need ~30 min of additional training to image BL2 samples!
- You must bring your PPE and secondary container for BL2 training! We will postpone your training if you do not bring these items.
Is my sample BL1 or BL2?
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BL2 (Risk Group 2) material contain potentially harmful agents such as oncogenic rDNA, human materials, human and non-human primate cell lines, human pathogens, or live rodents with human xenografts.
After fixation, most human and mouse tissues and cells are no longer pathogenic and therefore Risk Group 1. Fixed samples may be observed under standard BL1 laboratory safety conditions. Live non-human specimens such as mouse cell lines that do NOT contain human pathogens are also BL1.
Unfixed samples that include ANY human tissue or live human or primate primary cells or cell lines are considered Risk Group 2 agents and require BL2-level imaging. Imaging of live Risk Group 2 (BL2) samples is strictly controlled.
What the HEK?
What about cell lines like HeLa, MCF7, MEF, U2OS, OVCAR-3?
- Is it human? Is it alive? Then it requires BL2.
- Is it a rodent line? Then BL1 is fine.
- If you are unsure whether your cell line is of human or non-primate origin, search for it on ATCC.
Yes, imaging live HEK or HeLa cell lines requires BL2 and extra training! ALL live human or primate derived cell lines require BL2.
Still unsure? Please check with core personnel if you are unsure whether your sample is BL1 or BL2.
BL2+ specimens such as cells with active or live lentivirus are not permitted in the KIMCF.
Elevated BL2 Operation Requirements {#BL2}
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BL2 specimens such as live human-derived cells require the temporary raising of a room to BL2 status with proper signage and decontamination at the end of your session. See below.
Pay attention to BL1 and BL2 signs outside of each microscope room.
Research Registration Requirements {#amend_brr}
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Secondary transport containers are one of the most important aspects of BL2 operation. They should contain any spills while transporting your sample to, from, or within the core. The lids must stay on and the container must remain sealed if it is dropped! The secondary container must be liquid tight, have a latching lid, and tough hinges that will not break. You can find airtight plastic food storage containers with latching lids at many stores. Pelican also makes great safe secondary containers in their Micro Case series.
Do NOT use styrofoam containers or pipette tip boxes as secondary containers.
Sample Requirements
- Samples should be prepared and sealed in the user's home lab.
- Well-plates should be sealed with adhesive film and lids replaced. Apply parafilm to secure lids in place.
- Sample liquid volumes for BL2-level work shall be less than 0.5 mL per sample well, totaling to a maximum of 5.0 mL per sample plate.
- If preparing slides, coverslips should be sealed in place using nail polish or wax sealant such as VALAP.
- Consult with core personnel for alternate imaging chambers.
- All specimens MUST be transported to, from, and within the core in approved secondary containers. Well plates must be carried in a container with a locking lid. Slides may be placed within a 2nd container like a petri dish with lid which can be secured with parafilm during transport. For other cases, do not assume your secondary transport container is adequate. Always consult with core personnel for appropriate secondary containers prior to starting BL2-level work.
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