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Comparison of sample preparation methods for scanning electron microscopy( SEM) of leaf epicuticular waxes in Arabidopsis

XU Xiu-ping;GU Dan;FENG Min;State Key Laboratory of Systematic and Evolutionary Botany,Institute of Botany,the Chinese Academy of Sciences;Key Laboratory of Photobiology,Institute of Botany,the Chinese Academy of Sciences;  
Waxy layer of the plant epidermis is the first barrier to resistant environmental stimulation. Scanning electron microscopy( SEM) is a useful technique to observe the leaf epicuticular waxes of Arabidopsis. Different sample preparation methods for SEM observation were compared including critical point drying and freeze drying,FAA fixation and glutaraldehyde-osmium tetroxide double fixation. The effects of different organic solvents on wax structure were also studied. The optimal method to examine epicuticular waxes contains the following processes: glutaraldehyde-osmium tetroxide double fixation,dehydration of tert-butanol and ethanol gradient,and freeze drying.
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