Isolation of Protoplasts / Plant Cells
Arabidopsis thaliana belongs to the family of Brassicaceae (like cauliflower) and is a model organism for crop research. Since colza (Brassica napus) belongs to the same family Arabidopsis has become an interesting model for research on regenerative energy. The entire genetic code is known and its growth characteristics allow for investigation of several generations (generation time is about at 6 weeks) within short time.
Plant cells were treated with cellulase in order to remove the cell wall. Single cells in buffer were evaluated on the screen in phase contrast, bright field and fluorescent mode of the CellCelector microscope. Fluorescent plant cells can be detected automatically.
Fig. 1: Detailed image of the scan map. Fluorescent Arabidopsis thaliana leaf cells aside non-fluorescent cells.
The CellCelector facilitates the aspiration of a viable and intact cell, which is prerequisite for the downstream processes like RNA-analysis via single cell RT-PCR.
Fig. 2: Documentation of the picking of protoplasts in fluorescence channel by the CellCelector. Left: before picking; Right: after picking.
Fig. 3: Documentation of picking protoplasts in bright field illumination. Left: before picking; Right: after picking.
Fig. 4: RT-PCR for gdp2 of Arabidopsis thaliana leaf cells isolated by the CellCelector. Glycindecarboxylase (gdp) is part of the photorespiratory cycle in C4 plants.