Elements imaging software

Freshly confluent 24-well plates of HFFs or MEFs were infected with 100 parasites for each well and the plates were incubated at 37 °C without disturbing for 5 days. The total number of plaques in HFFs was observed using a Nikon TE2000 inverted microscope. To measure the parasite growth in MEFs, the area of each individual plaque was captured and analyzed using the above microscope equipped with Hamamatsu ORCA-ER digital camera, and NIS Elements Imaging Software, respectively. For each independent experiment, at least 40 plaques from technical duplicate wells were imaged.

Worms were picked into 5 µL of a 10-mM levamisol solution on microscope slides with 2% agarose pads. They were immediately covered with a coverslip, sealed with nail polish, and imaged using a Nikon TiE-C2 confocal microscope. The tps expression patterns were imaged with a 40x CFI Plan Apochromat (NA 1.25, water immersion) objective and the NIS Elements Imaging software (version 4.30.02). GFP was excited at 488 nm and the emission was collected with a 525-nm band pass filter. A z-series of images was taken (Z-step of 0.75 µm) using consistent imaging settings and a maximum intensity Z-projection was applied using the program Fiji [62 (link)]. The resulting Z-projections were stitched using Corel Photo-Paint^TM X5 (Version 15.0.0.486). Image tone was adjusted consistently for all images using the contrast enhancement tool (input value clipping 0-100). Compilation of images into panels was conducted in Microsoft PowerPoint.

Freshly confluent HFFs 24-well plates were used to determine relative parasite growth by plaque assay. On the day of infection, the media was replaced, and 250 freshly harvested parasites were added to each well. Plates were then left undisturbed for 6 days at 37°C 5% CO₂, after which plaque areas were imaged and measured. Plaque areas were captured and analyzed using a Nikon TE2000 inverted microscope equipped with Hamamatsu ORCA-ER digital camera and NIS Elements Imaging Software, respectively. For all experiments, at least 20–25 plaques from technical duplicate wells were imaged. For measurement of total parasite growth, a luciferase-based assay was performed [77 (link)].