Materials and Reagents

1. Mice. We perform most of our work in C57BL/6 mice due to the wide availability of transgenic and knockout strains on this genetic background. Slight differences in vendor-specific sub-strains exist, so it is preferable to obtain the mice from the same source throughout a given study. The majority of our studies use 9-12 week old females, since males have higher parasite loads and mortality with P. chabaudi infection in many mouse strains (Laroque et al., 2012).
   
2. Plasmodium parasites. Plasmodium parasites can be obtained from the Malaria Research and Reference Reagent Resource (MR4). The most commonly used rodent malaria strains include P. chabaudi AS (MRA-429; used to model uncomplicated malaria), P. berghei ANKA (MRA-311; fatal cerebral malaria), P. yoelii 17XNL (MRA-593; uncomplicated malaria; resolves late relative to P. chabaudi), and P. yoelii YM (MRA-755; lethal malaria without cerebral pathology).
   
3. Immersion oil Type A (Cargille, catalog number: 16482 )
   
4. Methanol (Thermo Fisher Scientific, catalog number: A412P4 )
   
5. Giemsa stain (Acros Organics, catalog number: 295591000 )
   
6. 1x PBS (Mediatech Inc., catalog number: 21-040 CV )
   
7. Alsever’s solution (MP Biomedicals, catalog number: 092801154 ) (see Recipes) (Note 1)
   

Equipment

1. 1 ml syringes (BD, catalog number: 309628 )
   
2. 25 g ⅝” needles (BD, catalog number: 305122 )
   
3. 25 mm x 75 mm x 1 mm unfrosted glass slides (Premiere, catalog number: 9101-E )
   
4. 25 mm x 75 mm x 1 mm frosted glass slides (Globe Scientific Inc., catalog number: 1324W )
   
5. Slide staining rack and chamber (Electron Microscopy Sciences, catalog number: 62543-06 and 62541-01 )
   
6. Glass cutter (optional) (General, catalog number: 06637383 )
   
7. 1.5 ml microfuge tubes (Thermo Fisher Scientific, catalog number: 50809238 )
   
8. Sharp surgical scissors
   
9. Brightfield microscope equipped with 100x objective
   
10. Lens paper (Thermo Fisher Scientific, catalog number: 11-995 )
    
11. Lamellar flow hood (optional; if working in a pathogen-free animal facility)
    
12. Scale with cup for weighing mice (OHAUS, catalog number: SB36853M )
    

Software

1. VersaCount (Kim and DeRisi, 2010)

Procedure