Spores do not have to be oval,a nd often they are round in monolayer
conditions.
Add non-ionic detergent, which will lyse amoebae but leave spores intact
and viable.  We use 1% Cemulsol, but this is no longer available.  triton
X-100 should do just as well.  Check with real spores and amoebae.

Rob


> Correction: "24 hours" should be "48 hours".
>
>
>
> -----Original Message-----
> From: Liao, Xin-hua (NIH/NIDDK) [F]
> Sent: Tuesday, March 31, 2009 12:55 PM
> To: [log in to unmask]
> Subject: [DICTY - 68] spore or stalk cells?
>
>
>
> Dear Dicty colleagues,
>
> I am developing sporulation assay in monolayer recently. Attached are
> the cells treated with 15mM 8-BR-cAMP for 24 hours with different
> density. At the beginning I thought all of the cells with bright
> periphery are spore cells. However, first, they are not elliptical;
> second, they do not express PSPA promoter driven GFP very well; third,
> sporulation efficiency looks too high if they are all spore cells.
> Alternatively, cAMP is also required for stalk differentiation. From the
> shape of the cells, I just can not tell whether they are spore cells,
> stalk cells or even undifferentiated cells.
>
> Any comments are welcome.
>
> Thanks,
>
> Xin-Hua Liao
>
>