For capsule staining I used over night bacteria culture and smear with 1% congo red . After that, dry the slide in room temperature and observe under light microscopy 100x magnification oil immersion . But I did not find good capsule layer around the cell surface. Can any one tell me better protocol for capsule staining with congo red ?
Do you fixate with 80% ethanol saturated with NaCl before congored use?
Enrico Marani actually I never used 80% ethanol saturated NACl for fixate. I used India INK and CV but result is not good. I used congo red but still i did not find capsule layer around the cell. However, can you explain saturated NACl with 80% ethanol protocol ? example 100 ml ethanol with 1 gram NACl ?
Matthew Ibbs can you tell me manvels solution protocol because I could not find their protocol. They just indicate 10% FeCl3 and 5% phenol, acetic acid like that.
For all classical stainings and solutions look into:
Romeis B (1948 there are more recent editions) Mikroskopische Technik Muenchen, Leibniz Verlag. and
G.L. Humason Animal Tissue techniques 3rd ed (1972) Freeman and Company San Francisko.
1% aqueous sodium hydroxide 0.5 ml add to 80% ethanol saturated with Na Cl 50.ml mix and use within 15 min, in other words make it fresh. Treat for 10-20 min.
Enrico Marani
I follow your protocol . Firstly, I made 50 ml of 1% aqueous NaOH with 80% etoH with Nacl . Then over night bacteria culture put into glass slide and dry with air. Then I put the solution into glass and smeared and dry. After that I used 1% congo red and dry. Observe into light microscopy. But I did not find CPS layer in my bacteria sample. Can you check.
You are correct, don't see it either. Dit you fixate after bacteria culture was dried on glass. Unclear in your description. Marani
Sorry I am not convinced that the method I recommended works looking to the results. There is some red at the cell periphery, but should not built results on it.
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