Exercise 13.7 - Isolation of Fruiting Bodies without slugs
- 24 suspension cultures of D. discoideum
- Clinical centrifuge and tubes
- Black Millipore filters (AABP47SO)
- Cellulose pads for Millipore filters
- LPS buffer
- 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).
- Apply the cells at a density of 6 X 10
cells per cm
to a water washed black Millipore filter supported by two cellulose
absorbent pads previously saturated with LPS Buffer.
- Allow the cells to develop in the dark at 22° C in a
humidity saturated chamber.
- 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 -- firstname.lastname@example.org