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How can i fix cultured cells on glass slides for insitu hybridization


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#1 NovelProtein



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Posted 19 August 2013 - 07:34 AM

Hello friends,


We have ordered mRNA--probe for our new protein that we are working on. Company people have provided positive and negative control slides (Human cell lines expressing RNApol2 n Dap). However, we have tried our probe over human "tissues" of our interest. Now parallel to this, we have endothelial cell lines which express our protein in high amount which we want to take as relative positive control.


So my question is....how can i fix our endothelial cell lines on glass slide so that i can perform mRNA based insitu hybridization over it and later can compare it with the human tissue sections. (its obvious that RNA's free condition is necessary). 


Apart, if anyone can suggest an alternative to use our probe in different manner than also please suggest (i.e if at all its possible to do it other way...than that on glass slide). 


Thanks in advance.



#2 bob1


    Thelymitra pulchella

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Posted 19 August 2013 - 01:04 PM

Check out cytospin protocols and have a talk to a histology lab about the best slides for keeping cells attached.  I suspect you will probably need specially coated slides like amino-silane coatings.

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