Plan apochromat 63x 1.40 oil m27

Intracellular tachyzoites grown in cover slips were fixed using cold methanol 100% for 8 min, washed with PBS and blocked with 1% BSA. Primary antibody αrH2B.Z (rabbit), αrSag1 (mouse), αrHSP90 (rabbit), αrROP5 (rabbit), αrROP18 (rabbit) produced in the laboratory and αc-Myc (rabbit, abCam ab9106 or mouse, Santa Cruz sc-42), αAcTubulin (mouse, Millipore MABT868) were incubated for 1 h at room temperature. After several washes with PBS cover slips were incubated with secondary antibodies Alexa fluor goat anti-mouse 488 and anti-rabbit 594 (Invitrogen). DAPI was used to stain nuclei. Axio Imager.M2 Microscope Carl Zeiss (Germany) with objective Plan-Apochromat 63x/1.40 Oil M27 and Video digital camera Zeiss 503 monochromatic 2.8 megapixeles, was used. Image J 1.53q (Fiji) software was used to process images.

Single layer fluorescence images were taken with a Zeiss AxioCam MRm (1,3883 x 1,040 pixels) at a Zeiss Axio Observer Z1 fluorescence microscope with Plan-Apochromat 63x/1.40 Oil M27 lens, 49 DAPI filter (Hoechst33342 detection), 78 HE ms CFP/YFP filter (γ-H2.AX AF-647 detection) and a transmission grid VH ‘ApoTome’ (Carl Zeiss, Oberkochen, Germany). Images were taken with three fourth of the maximum intensity without overexposure. The pictures were saved as 16-bit multi-channel Carl Zeiss Image files with no further editing. At least 80 nuclei per cell line per experiment were analyzed by counting γH2A.X foci. N = 3 independent experiments were performed and used for the analysis.

To evaluate the coverage and smoothness of the lipid layer adsorbed directly onto the device surface, confocal fluorescence microscopy experiments were performed by tagging the lipid solution with 0.1 mol% Texas Red^TM  DHPE dye. The morphology of lipid films on the sensor was monitored using a Zeiss LSM 800 confocal scanning laser microscope (Carl Zeiss AG, Germany) and a Plan-Apochromat 63X/1.40 Oil M27 objective lens. Supplementary Fig. 2 shows that the lipid coverage is excellent with virtually no defects detected in a micrometer lengthscale.