[Histonet] coverslipping question

Gautier, Nicole M. ngauti <@t> lsuhsc.edu
Tue Nov 12 08:32:24 CST 2013

My lab has been having a problem with "specks" appearing on our slides after they come out of Histoclear.  Our protocol is to dehydrate in 2 minutes each of 70%, 80%, 95%, and 100% ethanol before 2 minutes 2 times in Histoclear.  The slides are perfectly clear when they come out of the ethanols, but not when they come out of the Histoclear.
Since it's not a problem we had when we first started, I tried changing out the Histoclear, but the problem remained.  At the end of last week, I changed out everything - all glassware, all ethanols, all Histoclear.  The problem went away and the 24 slides looked fine.  Today, the slides are looking speckled again.
So I guess my questions are: What could be causing the speckled appearance of the slides? and  How often should I have to change the ethanols and Histoclear?  I seem to remember years ago when I did this that we only changed the solutions monthly...

Nicky Gautier
Research Associate

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