c4zhang at scimail.uwaterloo.ca
Sat Oct 25 18:28:29 EST 1997
The following is the information kindly provided by Melissa to my
question about vacuum infiltration in Arabidopsis fixation.
We have successfully used Karnovsky's fixative for all stages of
Arabidopsis pollen, from microsporocytes to mature pollen grains. The
recipe is: 4% paraformaldehyde, 3% gluteraldehyde in 0.05M potassium
phosphate buffer, pH 7.0. If you don't have experience making this type
of fixative, let me know, and I'll send you the detailed protocol. It is
difficult to get fixative to penetrate anthers and pollen, so we
usually add 0.01% Tween 20, and always vacuum infiltrate for a long time
(sometimes up to half an hour). You can see some of the results
in Spielman et al. (1997) Development 124, pp. 2645-57.
Let me know if you would like more information.
More information about the Methods