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- 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.
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- 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.
Sample Manipulation
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Area
- The fume hood in 76-280 may be used for brief manipulation of samples, including adding reagents or pipetting from well-to-well.
- Use pipettes and tip waste container kept within the fume hood. Do not remove. If you need additional equipment, please arrange with core staff.
- As with sample prepared in your home lab, samples must be transported within the core facility in a secondary container. Place your plate back in its secondary container to carry it to the imaging room!
- As with microscope rooms, a BL2 sign must be placed on the fume hood while it contains BL2 material.
- Use a fresh square of blotting paper for sample manipulation.
- Use the sink next to the fume hood for any emergencies. We will keep a supply of bleach under the sink in 76-280 and large buckets for sample decontamination above the sink.
- Small spills within or near the fume hood will be blotted and disinfected with 10% bleach using the spray bottle within the fume hood. Contact for at least 10 minutes, and dispose of any waste in the biological waste containers.
- When done, spray the blotting paper with 10% bleach, let sit for 2 minutes, and dispose in the biological waste container. Then spray the work surface of the fume hood and pipettes with 10% bleach, let sit for 2 minutes, wipe clean with a paper towel. Spray again with 70% ethanol and wipe clean to remove residual bleach.
- Dispose of sharps and pipette tips in the sharps box in the hood. Dispose of larger solid waste (paper towels, blotting paper) in the biological waste container.
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