In vitro quantitation of Theileria parva sporozoites for use - TropMed ...

Appendices 153
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Add 150 µl medium in each well using a multichannel pipette (Use a fresh set of tips for each
column). Pipette-mix about 3 times at each step.
Label the plate
Transfer to CO 2 incubator at 37°C (record time)
Leave undisturbed for 10 days
Day 10
Destroy fungi contaminated cells with 10N NaOH.
Check cell cultures for clumps
Make cyto-centrifuge smears: mount slides with a filter card, centrifuge 50 to 100 µl at 215 g
for 5 min
Stain with 12.5% Giemsa’s stain in Sörensen's phosphate buffer (0.008 M, pH 7.2)
Score cultures for the presence or the absence of schizonts