[Histonet] re vacuoles in nuclei of PT Link treated tissue

[Steven Weston]

Hi everyone,

I have recently encountered an unusual result in our immunostaining.

We have been bvery successfully using Dako's PT link with the High PH soln to dewax and heat retrieve some of our immuno samples. In conjunction with my colleagues from the local Hospital Anat Path dept we noticed a strange but prominent vacuolisation showing in the Nuclei (and less so in cytoplasm) in our treated tissue after staining with Haematoxylin. Here is the interesting thing, when we manually dewax the same type of sample and then through the high PH, PT link the vacuoles no longer appear. This was noticed in both laboratories. the first thought was that it had something to do with the wax. But we both use different waxes (precision cut and paraplast). Clearly it is something to do with the wax removal process but coming up with an explanation is challenging. Any ideas? Has anyone else noticed this phenomenon ?

regards

steve weston
lab manager
resp research group
MRI