Tecnai g2 t12 spirit biotwin microscope

TEM analysis was performed with Tecnai G2 T12 Spirit BioTwin microscope (FEI Company, Hillsboro, OR, USA). Before the analysis, the isolates (5 μL) were deposed on the formvar support on a copper mesh (200 mesh, Agar Scientific, Stansted, UK), contrasted with a 1% m/v uranyl acetate, and left for drying.

TEM imaging was performed according to the procedure described in [17 (link)]. The isolates (5 µL) were deposed on the formvar support on copper mesh (200 mesh, Agar Scientific, Stansted, UK). After solvent evaporation, the samples were contrasted with a 1% uranyl acetate and left for drying. The preparation was investigated with the use of the Tecnai G2 T12 Spirit BioTwin microscope (FEI Company, Hillsboro, OR, USA).

P. zantedeschiae 9M strain grown on agar media as well as MVs samples of P. zantedeschiae 9M wild-type and GFP-tagged strains were analysed with transmission electron microscopy (TEM). Bacteria were grown on the MH agar supplemented with 10% sucrose or the CVP agar plates for 48 h at 28 °C. MVs were obtained from bacteria grown in the M63 medium supplemented with 0.2% glycerol and 0.4% PGA with the method described in this manuscript (Section 4.5). TEM imaging was performed according to the procedure described by [27 (link)]. Briefly, 5 µL of the samples were deposed on the formvar support on a copper mesh (200 mesh, Agar Scientific, Stansted, UK). After solvent evaporation, the sample was contrasted with a 1% uranyl acetate and left for drying. The preparation was investigated with the use of the Tecnai G2 T12 Spirit BioTwin microscope (FEI Company, Hillsboro, OR, USA).
The MVs samples of P. zantedeschiae 9M wild-type and GFP-tagged strains were also analysed by confocal fluorescence microscopy. Vesicles were placed on a glass slide, sealed with a coverslip, and examined using the confocal laser scanning microscope (Leica SP8X, Germany) with a 100× oil immersion lens.