Manipulation of Ovaries/Ovules and Clearance and Isolation of Embryo Sacs of Impatiens glandulifera and Nicotiana tabacum

The objective of this research was to develop techniques to clear and/or isolate embryo sacs of Impatiens glandulifera and Nicotiana tabacum, to facilitate microscopic examination for subsequent experimentation on in vitro fertilization of flowering plants. Two techniques were applied: The clearing and enzymic maceration. Both of the 2 adopted techniques were effective in revealing embryo sacs of the studied plant species. A fixative prepared from propionic acid, formaldehyde, ethanol (50%) in the ratio 5:5:90 was used. The clearing solution was prepared from lactic acid (85%), chloral hydrate, phenol, clove oil and xylene in the ratio 2:2:2:2:1 by weight. Both the clearing and the enzymic maceration techniques proved effective in revealing embryo sacs of I. glandulifera and N. tabacum