There
are many methods of tissue sample fixation: from
picric acid to bouins solution to paraformaldehyde.
The following fixation protocol describes the
use of 4% paraformaldehyde (PFA).
note:
paraformaldehyde is toxic - do not inhale the
powder; work under a fume hood
1.
dissolve 4 g PFA in 50ml dH2O
2. add 2 drops 10N NaOH
3. add a stir magnet; stir and heat to 60 C -
until solution is clear
4. add 50ml 2x PBS
5. adjust pH to 7.3
6. if working with cryosections, immerse slides
with non-fixed tissue sections into PFA for 45
minutes
7. if preserving a whole tissue sample, immerse
sample into PFA solution (the solution should
be approximately ten times the volume of the tissue
sample) overnight
note:
PFA should be made fresh as needed |
