Spinning disc 100x microscope

After fixation, cells were permeabilized with 0.5% TritonX for 15 minutes. Primary antibodies were incubated for 1hour and washed twice with PBS. Secondary antibodies were incubated for 1 hour washed twice with PBS and cells resuspended in PBS containing Hoechst for observation under the microscope. Images were either taken on a Zeiss 200M Axiovert widefield (63x) or Nikon spinning disc (100x) microscope. Image processing was performed with ImageJ. Rabbit anti Trap: 1/100 (P. Sinnis), rabbit anti GFP for IFA 1/200 (abfinity), mouse anti GFP for western blot: 1/1000 (4mg/ml Roche). Goat anti mouse 594 1/1000 (Invitrogen 2mg/ml), goat anti rabbit 594 1/1000 (Invitrogen 2mg/ml), goat anti mouse HRP 1/10000 (GE Healthcare).

After fixation, cells were permeabilized with 0.5% TritonX for 10 minutes. For visualization of the red blood cell membrane cells were incubated with anti-Ter119::Alexa488 (Biolegend 0.5mg/ml; ¹/₁₀₀₀) antibody for 1h, washed twice with PBS, and resuspended in PBS containing Hoechst for observation under the microscope. For visualization of the parasitophorous vacuole (PV) membrane cells were incubated with anti-SEP1, followed by secondary anti-rabbit:: Alexa594 (Invitrogen 2mg/ml; ¹/₁₀₀₀). Images were either taken on a Zeiss 200M Axiovert widefield (63x) or Nikon spinning disc (100x) microscope. Image processing was performed with ImageJ.