Jacqui Detmar wrote:
Hi all. Having a bit of an internal debate here, so I would like to get
the opinions of some of you in Histoland, please. Here are the
questions:
1. When fixing with 10% NBF, for how long should you fix and what
volume ratio of fixative:tissue should you use?
48 hours is a minimum according to R.D. Lillie, one week is better.
Really! Thickness of the tissue does not matter, formalin fixes slowly.
Of course, if you are doing immuno you may need to sacrifice some
fixation to retain antigenicity. Depends on the antigen.
10X the volume of the tissue.
2. At what temperature should one be fixing tissues?
Room temperature. There is more than ample scientific evidence that
fixing cold just slows down the already slow fixation process.
Regarding Histochoice:
I never use proprietary fixes, you don't know what is in them and the
manufacturer can change the formulation any time he/she wants.
1. For how long should you fix the tissue?
2. What volume ratio of fixative:tissue should you use?
3. How long can you store Histochoice-fixed tissues in 70%
ethanol?
I think that's about it. Thanks in advance,
Jacqui
Jacqui Detmar, Post-doctoral Fellow
Samuel Lunenfeld Research Institute
Mount Sinai Hospital
25 Orde Street, room 6-1001AJ
Toronto, ON M5T 3H7
Email: det...@lunenfeld.ca
Phone: 416-586-4800 x5607
Fax: 416-586-5993
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Geoff
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcaul...@umdnj.edu
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