3 replies to this topic

[cruz]

Hi dear all,

I'm new here , and need some helps!

Our lab is working on a quite dangerous virus. We want to stain cells after infected virus. Since we want to stain nuclei using PI (to see apoptosis). For safety problem, we need to kill virus and then do cytometry. But! we also will kill cells at the same time, and PI will stain all dead cells...
How can we kill the virus without killing cells?? Is there any alternatives to stain cells to see apoptosis?
I apologize if I didn't explane the situation very well.

regards

C

[Doki]

cruz, on May 7 2009, 11:48 PM, said:

Hi dear all,

I'm new here , and need some helps!

Our lab is working on a quite dangerous virus. We want to stain cells after infected virus. Since we want to stain nuclei using PI (to see apoptosis). For safety problem, we need to kill virus and then do cytometry. But! we also will kill cells at the same time, and PI will stain all dead cells...
How can we kill the virus without killing cells?? Is there any alternatives to stain cells to see apoptosis?
I apologize if I didn't explane the situation very well.

regards

C

I don't know about killing the deadly virus but about 'apoptosis', PI stains all the dead cells not just those that underwent apoptosis. Annexin V might be a better choice for detecting apoptosis.

[cruz]

Hi Nabi,

Thanks for your reply!
So, you mean Annexin V can utilize on fixed cells for apoptosis?

regards
Cruz

Nabi, on May 7 2009, 05:09 PM, said:

cruz, on May 7 2009, 11:48 PM, said:

Hi dear all,

I'm new here , and need some helps!

Our lab is working on a quite dangerous virus. We want to stain cells after infected virus. Since we want to stain nuclei using PI (to see apoptosis). For safety problem, we need to kill virus and then do cytometry. But! we also will kill cells at the same time, and PI will stain all dead cells...
How can we kill the virus without killing cells?? Is there any alternatives to stain cells to see apoptosis?
I apologize if I didn't explane the situation very well.

regards

C

I don't know about killing the deadly virus but about 'apoptosis', PI stains all the dead cells not just those that underwent apoptosis. Annexin V might be a better choice for detecting apoptosis.

[Doki]

cruz, on May 9 2009, 02:50 AM, said:

Hi Nabi,

Thanks for your reply!
So, you mean Annexin V can utilize on fixed cells for apoptosis?

regards
Cruz

No! Annexin V is not for fixed cells. But, I think it should be possible to stain and then fix. TUNEL might be for the fixed apoptotic cells. I have no more idea than this I am afraid.