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vacuolation in MDCK cells


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#1 miraclestrain

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Posted 11 September 2009 - 06:22 PM

Hi there,

i am studying viral infection in MDCK cells. A 24hr culture with 3 hour infection period of influenza virus was fixed using acetone. Then subjected to immunoperoxidase staining, washed using PBStween, and finally counter stained using Meyers Hematoxylina and eosin, with acid- alcohol dip for 1 sec and alkaline wash for 20 secs.

The cells stain good. But I see vacuolation.

Can someone tell me what would be the reason

MS

#2 bob1

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Posted 13 September 2009 - 04:29 PM

Apoptosis possibly?

#3 miraclestrain

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Posted 13 September 2009 - 06:44 PM

Hi,

But why would Influenza virus cause cell apoptosis, just after 3 hours of incubation. I am sure it is not the virus, but what other chemical can cause apoptosis among the mentioned above?

Thanks
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MS

#4 bob1

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Posted 14 September 2009 - 05:14 PM

Ok, I interpreted your experiment as 3hr virus infection, then fixed 24 hours later, rather than the other way around. 3 hours is a bit short for all the cells to be showing apoptosis, and would be unusual unless the viral dose was massive.

Your cells haven't senesced? Also unlikely.

#5 miraclestrain

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Posted 17 September 2009 - 04:30 AM

Hi Bob,

Yeah exactly, the cells r not sensced as well. I discussed with my supervisor, he says it might be water which is seeping in after fixation, causing the vacuolation. It is not actually vacuolation, but globulation cos of water.. forming water bodies.

Wierd one right

MS




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