[Histonet] re cutting at 50 microns
Steven.Weston <@t> utas.edu.au
Mon Jan 16 15:49:08 CST 2012
The neurobiologists in our organisation regularly cut 50 micron sections on our leica cryotome. It is usually easy to do so particularly with brain tissue that has been prepared by fixing in parafomaldehyde and then incubating for a couple of days in 30% sucrose in buffer. The tissue can then be frozen directly in OCT (or similar) the cryostat using the fast freezing peizo area. If the cryostat doesn't adjust to 50 microns then the trick is to set it at 25microns and move the block down to just above the knife then back up again this will move the block forward 25 microns and then as you bring it down again it will move the second 25 microns giving you sections at 50. For brain tissue it should be cut at between –15 and –12 degrees C.
Hope that all makes sense.
Centre of Research Excellence for Chronic Respiratory Disease.
Menzies Research Institute
More information about the Histonet