Is there anyone has an experience in plaque assay using C6/36 cell line? I have problem in staining so I couldn't take photograph of the plaques forming in a whole well plate, however I could found plaques forming and pick them to be purified were always possible, but still staining seems impossible, crystal violet lead the cell lysis and the well become empty, I also try staining with agarose gel contains natural red, but seems not even helping because the gel become dark and the plaques become more invicible.
Thx Barbara,
I am the one who count without staining now. Since the plaque assay in c6/36 always reported to be unsuccesful, I think this is my achievement and I can only prove by picture of those stained wells. But the fail moment was there. Frustating ever lol.
I already tried what you did on Hela cells and resulting all the cells lysis I mean they following the rinse tap water and the wells became empty/almost no monolayer anymore. I've try staining at 7dpi and even 48hpi, the cell still lysis. less then 48hpi I couldnt see plaque clearly because plaques were still very small.
Neutral red Staining also I've done it. since it doesnt require to remove the gel I think it should be better. I did two ways, mixed with agarose gel tend to make the mnolayer become darken instead. I tried also direct staining by dilution of NR in pbs but a little bit different with that u mentioned. I will try the neutral red staining as exactly as u mention above.
After trying I will post again here..
I am glad that you pay more attention to my question Barbara. I appreciate it.
that paper I already have it^, thx for share anyway, the figure you might found out in that paper was exactly what I mean.. I also found some reports with no further explanation about unsuccesful of c6/36 plaque assay prior to culex flavivirus titration. Their solution was simple, change it to C7/10 monolayer and it works then. But unfortunately we dont have c7/10 in our lab.
I am still curious and Believe someday I will find a best way to stain this sensitif cell line.
I ended up reporting Plaque purification using C6/36 by direct observation. we can see through light behind the wall. I saw clear plaque and the amount were exactly depend on dilution. Thank you Barbara..
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