Parasites were fixed for IFA using 4% Paraformaldehyde and 0.03% glutaraldehyde, then permeabilized with 0.1% Triton-X100. Primary antibodies used were rat-anti-HA 3F10 (Roche, 1:100), mouse-anti-HA 6E2 (Cell Signaling Technology, 1:100), mouse-anti-V5 TCM5 (eBioscence, 1:100), rabbit-anti-V5 D3H8Q (Cell Signaling Technology, 1:100), and mouse-anti-PfPMV (from D. Goldberg 1:1). Secondary antibodies used were Alexa Fluor 488 and Alexa Fluor 546 (Life Technologies, 1:100). Cells were mounted to slides using ProLong Diamond with DAPI (Invitrogen). Fixed and stained cells were imaged using a DeltaVision II microscope system with an Olympus IX-71 inverted microscope. Images were collected as a Z-stack and deconvolved using SoftWorx, then displayed as a maximum intensity projection. Images were processed using Adobe Photoshop, with adjustments made to brightness and contrast for display purposes.
For imaging of parasite cultures using light microscopy, aliquots of culture were smeared onto glass slides and field-stained using Hema3 Fixative and Solutions (Fisher Healthcare), which is comparable to Wright-Giemsa staining. Slides were imaged using a Nikon Eclipse E400 microscope with a Nikon DS-L1-5M imaging camera. To measure parasite size, images were taken and parasites measured using ImageJ (NIH).
For imaging of parasite cultures using light microscopy, aliquots of culture were smeared onto glass slides and field-stained using Hema3 Fixative and Solutions (Fisher Healthcare), which is comparable to Wright-Giemsa staining. Slides were imaged using a Nikon Eclipse E400 microscope with a Nikon DS-L1-5M imaging camera. To measure parasite size, images were taken and parasites measured using ImageJ (NIH).