Athropomorphic mustard pollen tube aniline blue staining

, methods and
reagents
maternal liquid
1.Ice acetic acid
2.Ethanol
3.1 M NaOH (sodium hydroxide)
4.100 ml 1 M K2HPO4 (hydrophosphate d
5.100 ml 1 M KH2PO4 (potassium phosphate)
6.Fisher
7.Glyce
working solution
1.Ice acetic acid content of 10% ethanol solution (
tissue
use)
2.50 mM potassium phosphate solution: 4.17 ml 1 M hydrophosphate d'Potassium 0.83 ml 1 M Phosphate potassium d'Ahydrophosphate: 995 ml water, pH7.5
3.KPO4 solution (staining) with a blue content of 0.01%
4.Potassium phosphate solution (sealing agent) containing 50% glycelyceride is , instrument
1.Ultraviolet microscope
step three , step
1.Add 250 μl of
acetic acid to the
1.5 ml of centrifugal tube and soak the males in ice acetic acid for at least 1.5 hours. The tissue can be fixed overnight if needed.
2.Soaking the fixed tissue in a 1 M sodium hydroxide solution overnight is tissue softening.
3. Wash plant tissue three times with potassium phosphate. (Plant tissue is fragile at this stage)
4. The tissue was dyed for 5-10 minutes or up to 10 hours with 200 sl of aniline blue.
5. After dyeing, place the tissue on the slide and observe it under a UV microscope. If necessary, you can press the tablet to observe.
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