In article <320F5C23.3985 at cnsvax.albany.edu>,
dab at CNSVAX.ALBANY.EDU (Dmitry Belostotsky) wrote:
>Does anyone know a source of decolorized aniline blue or reference on
>how to prepare it? Thanks
Dear Dimitry, here is a recipe that I have used. Let the DAB solution stand
for a few days before using. Cheers, Adam
Decolourised Aniline Blue (DAB) for observation of pollen tube growth:
Mix 0.2 g of aniline blue and 4.6 g K3PO4 and store in the dark
Prepare a working solution by adding glycerol 1:1 after it has
Solution can be reused.
Preparation of flower samples for pollen tube observation :
1. Fix the plant material in acetic acid alcohol (1:3)
2. Remove the fixative from the samples and place the flowers in
water (rinse * 3).
3. Replace with 10 % sodium sulphite solution (w/v)
4. Autoclave 1210C for 15 mins.
5. Allow to cool and remove solution
6. Wash 2-3 times in dH20
7. Replace water with DAB/glycerol
8. Dissect the flower to reveal the ovaries
9. Mount and seal the flower using either nail polish, hot vaseline
or hot wax.
University of Adelaide/CSIRO Voice: +61 08 8303 8627
Division of Horticulture Fax : +61 08 8303 8601
Urrbrae, Adelaide Email: Adam.Vivian-Smith at adl.hort.csiro.au