A Smear Technic for the Cucurbitaceae

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Abstract
It is very difficult to make satisfactory smear preparations of species in the Cucurbitaceae by ordinary methods because, (a) the differentiation between chromosomes and cytoplasm is poor, (b) the pollen mother cells are held together in tissue-like masses, (c) the chromosomes are comparatively small and numerous. Special procedures have been devised to overcome these difficulties. Staminate buds selected at the proper stage of maturity are fixed for a period of 12 to 24 hours in a mixture composed of 3 parts of 100% ethyl alcohol and 1 part acetocarmine to which a small quantity of iron acetate has been added. After prefixation the material is rinsed in several changes of 100% alcohol. It can then be transferred directly to the slide for smearing, or hydrated to 70% alcohol for storage. For smearing the anthers are dissected out, the extraneous flower parts discarded, and the storage fluid removed. A drop of acetocarmine diluted to one-half strength with 45% glacial acetic acid is added, and the anthers are macerated into small pieces and smeared. The anther debris is removed, and the cover slip added. It is necessary to carry out the above operations with a low power binocular microscope. After heating the cover slip can be sealed with Pyseal for temporary storage.

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