Hi, I have been doing slide in situ hybridisation on wax sections (mouse embryos age e10.5) and every time I do it the tissue sections look damaged. Can anyone suggest a solution to improve the quality of my sections? Or suggest a reason for having tissue damaged sections?
Hi,
As suggested by Hussein, you can follow some protocols.
Here are some suggestions such as increase section tickness, decrease the fixation time, wash extensively with PBS after fixation, use Poly-L-Lysine coated slides, etc.,. If you have cryostat, I would suggest you to do in that.
First it comes to my mind to reduce the digestion-time. The longer the fixation the more digestion is needed. The shorter fixation the less digestion is needed.
Also the thickness of the slide has influence. The thicker the section the less sensitive for the digestion.
With heat-pretreatment one can also play around. The longer and more hot the less digestion is needed.
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