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Stigma characteristics and morphology can be useful in taxonomic and phylogenetic studies, indicate re- lationships in stigma function and receptivity, and be valuable in evaluating pollen-stigma interactions. Problematic is that in some taxa, copious stigmatic exudate can obscure the fine structural details of the stigmatic surface. Such is the case forCitrus,which has a wet stigma type on which abundant exudate inundates surface papillae. The components of stigmatic surface compounds are highly heterogeneous and include carbohydrates, proteins, lipids, glycoproteins, and phenolic compounds. This study evaluated the efficacy of several pre-fixation wash treatments on removing surface exudate to visualize the underlying stigmatic surface. Wash treatments included various buffer solutions, surfactants, dilute acids/bases, and solvents. Stigmas prepared using conventional fixation methods in glutaraldehyde had con- siderable accumulations of reticulate surface deposits with stigmatic cells obscured. Pre-fixation washes containing solventssuchasmethanol,chloroform,andethanolleftaccumulationsofincompletelyremovedexudateandcrystalline deposits. Alkalinewaterwashesproducedacrust-like depositonstigmasurfaces.Bufferwashesleft residuesofplaque- like deposits withperforatedareas.Incontrast,excellentremovalof stigmaticexudate wasobtainedwitha pre-fixation wash composedof0.2M Tris buffer, pH 7.2, containing 0.2% TritonX-100surfactant andallowed clearimagingof the stigma and surface papillae morphology. A central sinus and radially arranged openings on the stigmatic surface were clearly visible and shown for the first time using scanning electron microscopy (SEM).

Enhanced Visualization of the Fine Structure of the Stigmatic Surface of Citrus using Pre-fixation Washes