Willaertia magna De Jonckheere et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Alternate media: ATCC medium 711 inoculated with bacteria as above. (Note that ATCC medium 711 is a richer formula than ATCC medium 997 and will produce denser and faster bacterial growth. Excess bacterial growth can inhibit growth of amoebae.)
Storage and Culture Initiation
Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen. If liquid nitrogen storage facilities are not available, frozen ampoules may be stored at or below -70°C for approximately one week. Do not under any circumstance store frozen ampules at refrigerator freezer temperatures (generally -20°C). Storage of frozen material at this temperature will result in the death of the culture.
Note: a monoxenic amoeba culture can be established in this manner using any suitable bacterial food source.
DMSO, 1.5 ml
Page's Balanced Salt Solution (or similar), 8.5 ml
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Michel R, et al. Ultrastructure of the amoebo-flagellate Protonaegleria westphali. Parasitol. Res. 74: 23-29, 1987. PubMed: 3438290