| Reference |
Zavřel, T., Očenášová, P., Červený, J (2017): Phenotypic characterization of Synechocystis sp. PCC 6803 substrains reveals differences in sensitivity to abiotic stress. PLoS ONE 12(12): 1-21, DOI: 10.1371/journal.pone.0189130PubMed ID29216280
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| Method |
P.7 bottom paragraph: "Cell size was determined using ImageStream MkII imaging flow cytometer (Amnis Corp.,
Seattle, WA, USA). Right after harvesting from the photobioreactor, 500 μl of the culture suspension was centrifuged (4 000 g, 4 min, 25˚C), supernatant was discarded, pellet was resuspended in 0.25% glutaraldehyde solution and the samples were incubated for 10 min at laboratory temperature. The fixed cells were stored in -80˚C until further processing (up to 1
month in total). For further analysis, the samples were thawed on ice for 2 hours, and they
were kept at laboratory temperature for additional 30 min after thawing. During the cytometric analysis, only bright field images were collected. Gating of the measured populations
was applied to discriminate: a) focused objects (using combination of both RMS gradient and
Treshold Mask features of IDEAS1 software), and b) round objects (width/ length ratio
between 0.9–1.0). The imaging flow cytometer was calibrated with nonfluorescent microspheres (1–15 μm, Thermo Fisher Scientific, Waltham MA, USA) and the results were validated with light microscope Axio Imager 2 (Carl Zeiss, Oberkochen, DE)." |