Cell morphology was routinely checked using a Zeiss Axio Observer.Z1/7 microscope equipped with a LCI Plan-Neofluar 63x/1.3 water-immersion objective. Cell morphology was analyzed in bright-field mode. Pictures were processed with the Zen 2.3 lite (blue edition) software (Carl Zeiss Microscopy GmbH).
For cell size and viability measurements respective cultures were washed and diluted in PBS (KH2PO4 0.24 g L-1, Na2HPO4 2 H2O 1.8 g L-1, KCl 0.2 g L-1, NaCl 8 g L-1) followed by brief sonicatation. For viability measurements, cells were stained in the presence of 2mM propidium iodide (PI) before fluorescence (638 nm laser), forward and sideward scatter were measured at CytoFlex S flow cytometer (Beckman Coulter). Ethanol fixed cells were used as negative control. The size distribution of the cell population and the mean cell size was determined by calibration using a polystyrene particle size standard kit (SPHERO Flow Cytometry Size Standard Kit, Spherotech). Flow cytometry data was analyzed using CytExpert (version 2.3.0.84).
For cell size and viability measurements respective cultures were washed and diluted in PBS (KH2PO4 0.24 g L-1, Na2HPO4 2 H2O 1.8 g L-1, KCl 0.2 g L-1, NaCl 8 g L-1) followed by brief sonicatation. For viability measurements, cells were stained in the presence of 2mM propidium iodide (PI) before fluorescence (638 nm laser), forward and sideward scatter were measured at CytoFlex S flow cytometer (Beckman Coulter). Ethanol fixed cells were used as negative control. The size distribution of the cell population and the mean cell size was determined by calibration using a polystyrene particle size standard kit (SPHERO Flow Cytometry Size Standard Kit, Spherotech). Flow cytometry data was analyzed using CytExpert (version 2.3.0.84).