Accuri c6 software 264

The possible effect of the AC field on the membrane permeability of the cells was investigated by flow cytometry (FCM) using propidium iodide (PI, Life Technologies Europe B.V, Zug, Switzerland) stain. FCM analyses were performed using a BD Accuri C6 flow cytometer (BD Biosciences, San Jose, CA, USA) with an Accuri CSampler (BD Biosciences, San Jose, CA, USA). The number of cells and PI fluorescence emitted at 585 nm after excitation with a 488-nm argon laser were acquired and analyzed using the BD Accuri C6 Software 264.15 (BD Biosciences, San Jose, CA, USA). Data were collected to 10,000 events for each sample. C. reinhardtii and C. meneghiniana treated with 50 µM hydrogen peroxide (Life Technologies Europe B.V, Zug, Switzerland) were used as positive controls, whereas for Synechocystis sp., the cell suspension was heated at 70 °C for 20 min. For all phytoplankton species, the negative control corresponds to the cell suspension with no AC field treatment. PI at 7 µM was added and incubated for 30 min in negative and positive controls, as well as in all samples prior to FCM analysis.

Chlorophyll fluorescence of C. reinhardtii was followed in parallel with FCM. FCM analyses were performed using a BD Accuri C6 flow cytometer (BD Biosciences, San Jose, CA, USA) with an Accuri CSampler (BD Biosciences, San Jose, CA, USA). The data such as the number of counted cells and chlorophyll fluorescence from red channel (670 nm) when excited with 488 nm argon laser were acquired and analyzed using the BD Accuri C6 Software 264.15 (BD Biosciences, San Jose, CA, USA). Data were collected to 10,000 events for each sample after 30, 60, 90, and 120 min exposure to contaminants.