I would like to study the karyotype of some plants species in arid regions, but I do not know what is the best thechnique used.
I find it easier to karyotype with young anthers. Ideally these would be collected before turning yellow (which indicates that pollen maturation has occurred). Select anthers that are still white/opaque. Stain these with acetocarmine by adding one drop and crushing the anthers completely. An additional drop may be necessary to spread the anthers and prevent drying while macerating. You should see them turn dark and any PMCs will be apparent as small specks (rather than clumps) as they darken. After staining 5-10 minutes, transfer to a clean portion of the slide. Only soak up the "liquid portion" with tweezers, which will gather cells but leave the anther wall behind. Add a drop of Hoyer's mounting medium and blend. Apply a cover slip and press between a paper towel holding two opposite corners of the cover slip. Slide air bubbles out with you needle tip. Use a little Hoyer's to seal the cover slip and you are ready to scan!
Hope this helps!
I find it easier to karyotype with young anthers. Ideally these would be collected before turning yellow (which indicates that pollen maturation has occurred). Select anthers that are still white/opaque. Stain these with acetocarmine by adding one drop and crushing the anthers completely. An additional drop may be necessary to spread the anthers and prevent drying while macerating. You should see them turn dark and any PMCs will be apparent as small specks (rather than clumps) as they darken. After staining 5-10 minutes, transfer to a clean portion of the slide. Only soak up the "liquid portion" with tweezers, which will gather cells but leave the anther wall behind. Add a drop of Hoyer's mounting medium and blend. Apply a cover slip and press between a paper towel holding two opposite corners of the cover slip. Slide air bubbles out with you needle tip. Use a little Hoyer's to seal the cover slip and you are ready to scan!
Hope this helps!
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