[Histonet] Help! NBF fixed tissue and subsequent first step for dehydration.

From:Lorraine Rolston

I would appreciate any advice concerning possible detrimental effects that can be caused to the morphology of NBF fixed tissue when placed directly into 70%ethanol from NBF. This tissue (pancreas, spleen, kidney)is  for research purposes (not clinical diagnosis) and  good preservation of cell morphology is of extreme importance.
  One of the research groups that uses our processing facility brings the tissues to us immersed in 70% ethanol ready for processing.They are then embedded and cut for H&E staining.
  Before their tissues are bought to the lab at the 70% ethanol step, they are washed in water after fixation,  then immersed in 50%alcohol.  At some stage this protocol has been changed and the 50%ethanol step has been   omitted. Coinciding with this step being discontinued has come the appearance of bad cracking of the tissue.The head of the research group now feels that the changes to the morphology of these samples is caused by the jump from the NBF fixative to 70%ethanol.
  The pancreas samples are processed separately using  chloroform for clearing.
  I have yet to view the slides but may be able to submit a picture(if given permission) when I do.
  Any helpful advice or ideas would be very much appreciated.
  Thanks in advance, Lorraine.

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