All cultures should be considered potentially hazardous and should be opened by individuals trained in microbiological techniques working in facilities with containment requirements appropriate for the biosafety level of the cultures. ATCC recommends that the handling or opening of glass ampoules be performed in a biological safety cabinet. If this is not possible, wear protective clothing, gloves, a face shield or safety goggles, and hold the vial away from your body. Ensure that all empty vials are sterilized before disposal.
Opening Double-Vial Preparations (see diagram below)
- Heat the tip of the outer vial in a flame.
- Add a few drops of water on the hot tip to crack the glass.
- Strike the end of the vial with a file or pencil to remove the tip.
- Remove the insulation and inner vial with sterile forceps. Gently raise the cotton plug.
Opening Single-Vial Preparations (see diagram below)
- To recover the cell suspension from the glass ampoule, score the neck of the ampoule with a small, sterile file.
- Disinfect the outside of the ampoule with freshly prepared 70% ethanol or dip it into a beaker of freshly prepared 70% ethanol.
- Wrap the ampoule within several folds of a sterile towel or gauze to dry residual ethanol.
- Working in a laminar flow hood, hold the vial upright and snap open the vial. Ensure that your gauze does not become too wet with ethanol, or alcohol could be sucked into the culture when the vacuum is broken. Rehydrate the material immediately