[Histonet] Cryojane tape-transfer and hydration of sections
amaluenda at svi.edu.au
Tue Jun 9 19:19:34 CDT 2015
I was wondering if there is anyone out there who is currently using the CryoJane Tape-transfer system for bone sections. I have been trying to get good sections for mice tibia and femur and often find myself with problems.
I spent a good time trying to improve the quality of sections (tried different temperatures, thicknesses and speed as well as 1x vs 4x coated slides). The sections improved (although not perfect), but I was planning to move on for a staining trial. Currently, the sections are taken at 5um, in a 4x coated slide, at -26oC. However, now I get myself with a problem when hydrating the sections. At the moment, once the sections are taken, they are kept in -20oC (for short-storage). They are then taken out and left at RT for 30 min before hydration with PBS for another 30 min. I have noticed that it seems as if the OCT around the sections doesn't completely dissolve. I have already tried PBS vs dH2O and played around with times (from 5 min to overnight) with no differences.
Has anyone had this happening before? Can it be because of the 4x coat? Is there anything I can do to? And would this be a problem for immunofluorescence?
Any advice would be much appreciated!
Thanks in advance,
Stem Cell Regulation Unit
More information about the Histonet