showing CPE by Giemsa staining - (Mar/12/2007 )

Hi;

İs anyone knows Giemsa staining method for showing CPE in cell culture flask?

Best regars.

may this help you!!!!!!!!!!!!!








Giemsa stain


i) Preparations of tissue culture supernatant, smears or impressions of the kidney, liver, and spleen should be prepared, air-dried, and fixed for 5 minutes in absolute methanol.


ii) Immerse slides in a working solution of Giemsa stain for 30 minutes.


Stock solution: 0.4 (w/v) in buffered methanol solution, pH 6.9 (commercially available).


Working solution: diluted 1/10 in phosphate buffer pH 6.0 (0.074 M NaH2PO4, 0.009 M Na2HPO4).


iii) Destain with tap water.


iv) Observe slides under oil immersion. Tissue smears from infected organs show darkly stained pleiomorphic organisms occurring in coccoid or ring forms, frequently in pairs, with a diameter of 0.5-1.5 µm.