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Instruction for Reviving Freeze-dried Cultures

Manipulate following procedures as carefully and aseptically as possible, using a clean bench. Moreover, store the ampoules in the refrigerator when they are not opened immediately.

  1. Prepare liquid media, solid culture media and sterilized Pasteur pipettes. Letters on ampoules may be vanished by the sterilizing alcohol. Affix to the ampoules paper labels with the information necessary for identification.
  2. Score the central part of the ampoules with an ampoule cutter.
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  3. Sterilize the surface of the ampoule with absorbent cotton wetted with 70% ethanol.
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  4. Cover the ampoules with sterilized gauze. Then break the ampoules carefully at the scored part.
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  5. Add 0.1ml of liquid media or sterilized water to the ampoule and suspend the dried samples.
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  6. Aspirate the dissolved samples and drip it on a plate. Streak the suspension as illustrated below. Flame the loop after the 1st and 2nd streak.
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  7. Incubate the plate. It may take a double-period of general culture, since these cells are weak because of freeze-drying.