decolorized aniline blue

[viv000]

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
>Dmitry

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 
	overnight.

	Prepare a working solution by adding glycerol 1:1 after it has 	
decolourised.

	Solution can be reused.

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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.


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Adam Vivian-Smith
PhD Student
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
S.A., 5064
AUSTRALIA
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