Coolsnap hq2

For fluorescent microscopy analysis, cells were grown at 37˚C in accordance with the marker frequency analysis using glucose (2.0%) as the carbon source for cell resuspension. Individual cell membrane were stained with 0.4 μg/ml FM5-95 (N-(3-Trimethylammoniumpropyl)-4-(6-(4(Diethylamino)phenyl)hexatrienyl) Pyridinium Dibromide) from ThermoFisher Scientific. Cells were immobilized onto 1.2% agarose in 0.25x SMM base medium spread thinly onto microscope slides. Microscopy was carried out with Nikon Eclipse Ti equipped with Nikon DM 100x/1.40 Oil Ph3 objective, Photometrics CoolSnap HQ² cooled CCD camera and light source was Lambda LS (Sutter Instrument). Metamorph V.7.7 were used to acquire images. Cells were quantified using Fiji (Schindelin et al., 2012). At least three biological repeats were performed for each experiment.

For fluorescent microscopy analysis, cells were grown at 37˚C in accordance with the marker frequency analysis using glucose (2.0%) as the carbon source for cell resuspension. Individual cell membrane were stained with 0.4 μg/ml FM5-95 (N-(3-Trimethylammoniumpropyl)-4-(6-(4(Diethylamino)phenyl)hexatrienyl) Pyridinium Dibromide) from ThermoFisher Scientific. Cells were immobilized onto 1.2% agarose in 0.25x SMM base medium spread thinly onto microscope slides. Microscopy was carried out with Nikon Eclipse Ti equipped with Nikon DM 100x/1.40 Oil Ph3 objective, Photometrics CoolSnap HQ² cooled CCD camera and light source was Lambda LS (Sutter Instrument). Metamorph V.7.7 were used to acquire images. Cells were quantified using Fiji (Schindelin et al., 2012). At least three biological repeats were performed for each experiment.