I cut fixed frozen coronal sections of brain. I cut at -20, warm on a slide warmer with nice looking results. However, at a certain level, I seem to have problems. At this level, the thalamus is separated from cortical grey matter by a substantial amount of white matter. Cutting a section results in a differential expansion - the grey matter on either side of the white matter expands more than the white matter itself, so ruffles of tissue form. I use subbed coverslips to collect these sections, and upon collection, the ruffles of tissue melt to the slide with bubbles underneath the tissue on either side of the white matter. At the very least, there are notable folds. In order to get around this, I have altered the following variables: knife angle, roll bar angle, chamber temperature, velocity of cut and more. I have also tried pressing the coverslips onto the sections, delicately attempting to paint the ruffles away, warming coverslips, using slides, warming the collected sections at a slower rate than on the slide warmer. I have even lightly exhaled on the section to give it more form, as a web site recommends. All to no avail.
If you have any hints, or have faced this problem before, could you please let me know.
rushmore at bu.edu
-------------- next part --------------
An HTML attachment was scrubbed...