I am working on EAN Model, to perform histology i need very good slices, but unfortunately i couldn't get proper slices, tissues get distorted when we stain, we have made all possible changes but couldn't proper tissue for histology. any body doing HE or Immunostaining of sciatic nereve could guide me, Thanks.
Hi Sebastian I am working in Neurology clinic with Dr. Marcus Muller. I am using this following prcedure for Parrafin embeding,
after geting out the nreve, Fixed into 4% PFA for 21 hours, then ash it with 1x PBS three times after every 10 minutes. then
15 minutes in 50% isopropanol
15 minutes 50% isopropanol
• 15 min in 70% isopropanol
• 15 min in 70% isopropanol
• 30 min in 90% isopropanol
• 1 hour in 96% isopropanol
• 1 hour in 96% isopropanol
• 1 hour in 100% isopropanol
• 1 hour in 100% isopropanol
• 3 h in 100% isopropanol
Then putting the nerve tissue into incubator Overnight in isopropanol - paraffin (1: 1) at 60 ° C.
then
2 h in paraffin I
• 2 h in paraffin II
in paraffin III for overnight, then making blocks and cutting, but after cutting tissue looks destroyed and artifacts appeared in slices which is not solved yet.
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