Dear Dr. Xin-Hua Liao,

The photos are V12 or  NC4-derived cells?
Anyway, you can check spores and stalk cells by adding TirtonX-100  
(final ~0.1-0.5%) (as Rob says) or staining cell walls with Calcofuor  
(~0.1%).  And if you add neutral red (eg. final 0.0005% or less) into  
culture media from the beginning, you can observe presalk cells that  
have big vacuoles stained well with neutral red.

Best wishes,
Yuzuru Kubohara
> 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
>
> <01.jpg><03.jpg><09.jpg>


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Yuzuru Kubohara Ph.D.
Gunma University
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