Live acquisition software

Fluorescence imaging was performed on a IMIC digital microscope (FEI, Type 4001) using the Polychrome V light source, an Orca-R² camera controller from Hamamatsu (C10600) and Live Acquisition software (FEI, version 2.6.0.14). Image analysis was performed using Offline Analysis software (FEI). Bright-field imaging was performed on a Nikon E600 microscope equipped with Nikon objectives (Plan Fluor ELWD 20x/0.45, Plan Apo 40x/1.0 Oil and 60x/1.40 Oil) using a Digital Sight DS-UE camera controller and DS-Ri1 camera (both Nikon). Image analysis was performed using Nikon software NIS Elements 4.0.

Fluorescence imaging was performed on an iMIC digital microscope (FEI, Type 4001) using the Polychrome V light source, an Orca-R2 camera controller from Hamamatsu (C10600), and Live Acquisition software (FEI, version 2.6.0.14). Image analysis was performed using Offline Analysis software (FEI). Single-channel images are displayed in grayscale; merged images are displayed in false colors to avoid red-green combinations.

For morphometric analyses live parasites were cell surface-labelled with 3 mM AMCA-sulfo-NHS (sulfosuccinimidyl-7-amino-4-methylcoumarin-3-acetate, Thermo scientific, Pierce, Rockford) essentially as described in [24 (link)]. The cells were fixed in a final concentration of 4% w/v formaldehyde and 0.25% v/v glutaraldehyde in 0.1 M HEPES buffer over night at 4°C. Fluorescent microscopy was done using the automated iMIC microscope equipped with a Pike camera (PCO AG, Kelheim, Germany). The iMIC was controlled by Live Acquisition software (FEI Munich, Germany). 3D models of fixed cells were computed from deconvolved high-resolution 3D image stacks image stacks (z = 100 nm) using the Huygens Essential Image processing software v4.3 (SVI, Hilversum, Netherlands) and the Amira software v5.6.0 (FEI). An edge detection filter (Sobel) was applied and volume models were produced in Amira (Voltex display). Cells were selected for surface modeling (Isosurface display) and completed with the function ′pointwrap′. Flagella were traced using the volume model and Amira′s filament editor.