Methods and Techniques Discussion => Apoptosis or Cell Death => Topic started by: voland on July 23, 2008, 08:35:03 AM

Title: TUNEL AND DAPI
Post by: voland on July 23, 2008, 08:35:03 AM
I am staining rat tissue with TUNEL and DAPI and many cells that are TUNEL positive dont stain with DAPI. How do you interpret this.
Title: Re: TUNEL AND DAPI
Post by: excalibur on July 25, 2008, 09:48:51 AM
The cells are picking up the TUNEL and the DAPI has no where in that cell to connect?
Title: Re: TUNEL AND DAPI
Post by: voland on July 25, 2008, 10:57:44 AM
That's a possibility but is this a known effect/fact. Most of the reports show nice pictures with TUNEL-DAPI doucle staining.
Title: Re: TUNEL AND DAPI
Post by: richard03 on October 04, 2008, 05:32:24 PM
DAPI is not specific apoptosis marker. It just stains nuclei of cells and you identify the apoptotic cells based on its morphology. In later stage of apoptosis, all nuclei break apart and DAPI may not be able to stain them anymore. So TUNEL should be more reliable marker for apoptosis.
Title: Re: TUNEL AND DAPI
Post by: wormlover on October 27, 2008, 03:04:58 PM
Voland,

I am having the same problem with my TUNEL/DAPI staining in worm tissues.  Did you ever figure out an explanation?  Is it just that the cells are in the later stages of apoptosis and the DNA is too degraded?  I just wanted to confirm that my TUNEL positives are actually nuclei....

sarah
Title: Re: TUNEL AND DAPI
Post by: voland on November 12, 2008, 06:39:53 PM
Hi Sarah,

I dont know what the problem is. Maye your explanation is good but according to some apoptosis experts it should not be like that, DAPI should stain.