Fluorescence and scanning electron microscopy of citrus sinensis protoplasts

The technology of protoplasts isolation is an important tool for studies of plasmatic membrane biochemistry and physiology and cell wall regeneration. Protoplasts of “Caipira” sweet orange (Citrus sinensis L. Osbeck) were enzymatically isolated from cells in suspension and subsequently cultured in liquid medium. This study aimed to evaluate the viability of protoplasts of “Caipira” sweet orange freshly isolated by the Fluorescein Diacetate (FDA) stain, which showed 95% of fluorescing protoplasts. Through the technique of scanning electron microscopy, it was observed protoplasts of “Caipira” sweet orange freshly isolated. It was used Calcofluor White (CFW) fluorescent stain to visualize the cell wall regenerated protoplasts after 8 days following culture of the above-mentioned variety, wherein was observed the cell wall through a bright blue ring intense fluorescence..