A Rapid Method for Isolating Single Cells from Apple Flesh

The identification, separation and analysis of individual living cells can be used to analyze the heterogeneity and operation mechanism of living systems. The study of fruit development is based on the extraction of active single cells. In this study, we investigated the effects of different enzymes and enzymatic hydrolysis times on the extraction of single cells from the 'Fuji' apple (Malus × domestica Borkh. 'Fuji'). The results showed that the extraction of single cells in apple flesh was a suitable method when 0.1% macerozyme was used and the enzymolysis time was 0.5 h. Fluorescent brightening agent VBL staining showed that the cell wall was intact, while fluorescein diacetate FDA and azo dye Evans blue staining indicated that the extracted single cells were active. The extracted single cells could be further used as materials for protoplast extraction.