[Histonet] DAKO FISH pretreatment buffer (MES)

Kim Merriam kmerriam2003 <@t> yahoo.com
Mon Jun 23 08:59:43 CDT 2008


Hello,
This might seem to be a strange question, but I have been working up my FFPE FISH procedure with the DAKO FISH Histology Kit (I have had some nice preliminary results).  I have been noticing a strange greasy film on my slides that seems to be occuring during the heat pretreatment step (DAKO pretreatment buffer, which is MES; a very long name and a buffer I have not used until now); there is also a greasy film on the top of the buffer solution.
Is this common?  Can it hurt my slides?  I did manage to remove it by dehydrating my slides through 95, 100 and then clearing in xylene.  I then rehydrated them and continued my FISH procedure, I am in the middle of staining, so I won't know if it worked until tomorrow.  I am not even sure if I should continue and add my probe, which is hard to come by.
You might be thinking that there may still be wax on my slides, I thought of that too, but I deparaffinized the same as I always do for IHC, with 3 (3-minute) changes in xylene, 2 (3-minute) changes of 100% and then a change each of 95% and 80% before hydration in water.  I have never had this problem with my IHC staining or anything else, so I am inclined to think that it is the MES buffer that has some type of greasy component to it.
What does everyone think?
 Kim Merriam, MA, HT(ASCP)
Cambridge, MA


      


More information about the Histonet mailing list