Exercise 13.7 - Isolation of Fruiting Bodies without slugs

LEVEL III

Materials

Procedure 27

  1. Cultures grown for 24-30 hours (agar or suspension) are washed free of bacteria by several centrifugations in fresh, cold water (2 minutes @ 300 Xg each).

  2. Apply the cells at a density of 6 X 10^6 cells per cm^2 to a water washed black Millipore filter supported by two cellulose absorbent pads previously saturated with LPS Buffer.

  3. Allow the cells to develop in the dark at 22° C in a humidity saturated chamber.

  4. Periodically withdraw a sample of the amoeba and sketch the development of the aggregates. Note in particular the induction of synchronous fruiting without the formation of migrating slugs.

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Cell Biology Laboratory Manual
Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College,
St. Peter, MN 56082 -- cellab@gac.edu