free floating

[Annica Andersson]

Hi,
 I have a problem with free floating sections, I cut fresch froozen rat brain (no perfusion before sacrified) 40 um thich into etylenglycol/glycerol/PBS 1:1:2 and tissue broke almost dissolved the a tried another approach to section in to 2% pf and the same thing happened. Can it be because no perfusion?

[richard03]

If the tissue was fresh frozen, you must mount the sections directly onto slides, let them dry and then fix with formalin or acetone etc.

If you were doing  free floating sections, the tissue must be pre-fixed before frozen. In most of case like brain, it would be better to do perfusion and get better fix of the brain tissue.

Richard

[landras]

and if you cut the brain with a cryostat don't forget to keep the brains in 30% sucrose after the fix them, they should stay in sucrose until they reach the bottom of the tube.