See protocol on Chaos main page
- Use culture-treated dishes
- keep in Chaos saline at 22-24C in the dark
- Every 2-3 days
- change medium
- Split every 2-3 weeks, or when the debris becomes detrimental
- 7-day old Paramecium multimicronucleatum cultures at late log phase or early stationary phase (Fok and Allen, 1979).
- Remove growth medium:
- Filter cultures through a 3-piece glass filter funnel (Whatman)
http://www.whatman.com/GlassMicrofiberAccessories.aspx 1950-007 Filter Funnel, 3-piece, 70 mm, 115 ml reservoir 1/pk Fisher 09-877-3 $223.58
lined with a layer of GF/A glass microfibre filter (Whatman) 7.0cm dia. 1820-070
Fisher 09-874-16A Pack of 100 for $48.50
Wash and concentrate Paramecia in Chaos saline
- concentrate by allowing the medium to be filtered through
- collect concentrated Paramecium suspension in a beaker
Estimate Paramecium concentration
- 10 10uL drops
- count # Paramecium in each
- average x 100 = cells/mL
Feed 36 ± 3 Paramecia per amoeba.
Changing the medium
- Tilt the dish, let amoebae settle to the bottom
- Suck out the old medium
- Carefully squirt fresh medium onto the dish surface
- Repeat 2 or 3 times
- Check the culture
- Fast growing and healthy cultures have little cell debris (dead amoebae, cell fragments) and usually few or no lingering Paramecium left from the previous feeding.
- slow-growing or sick cultures have more cell debris and a large number of Paramecium left from the previous feeding.