Trophozoites of A. castellanii (ATTC
30234) were cultured in PYG 712 medium at room temperature in 75 mL
tissue culture bottles (Sarstedt, Germany), as described earlier.36 (link),37 (link) For each experiment, a PDMS pillar array was detached from the glass
slide and placed onto the bottom of a glass Petri dish (ibidi GmbH,
Germany). A 100 μL solution containing spherical particles,
200 μL of solution containing rod-shaped particles or no solution
was added to the dish, depending on the experiment. A. castellanii were detached from the cell culture
substrate by striking the container and shaking it vigorously. The
acanthamoebae were counted with a Neubauer hemocytometer, and 20 000
cells were incubated in the Petri dish in 1 mL PYG 712 medium for
15 h to ensure that the amoeba phagocytosed enough particles and that
the medium sufficiently wet the hydrophobic PDMS micropattern. In
total, 33 movies of A. castellanii migrating
on the samples were collected using a 60× oil immersion objective
(UPlanSApo 60× Oil, Olympus, Japan) on an inverted microscope
(IX-81, Olympus, Japan) and a digital camera (Hamamatsu C9300, Hamamatsu,
Japan) at 10 fps.
30234) were cultured in PYG 712 medium at room temperature in 75 mL
tissue culture bottles (Sarstedt, Germany), as described earlier.36 (link),37 (link) For each experiment, a PDMS pillar array was detached from the glass
slide and placed onto the bottom of a glass Petri dish (ibidi GmbH,
Germany). A 100 μL solution containing spherical particles,
200 μL of solution containing rod-shaped particles or no solution
was added to the dish, depending on the experiment. A. castellanii were detached from the cell culture
substrate by striking the container and shaking it vigorously. The
acanthamoebae were counted with a Neubauer hemocytometer, and 20 000
cells were incubated in the Petri dish in 1 mL PYG 712 medium for
15 h to ensure that the amoeba phagocytosed enough particles and that
the medium sufficiently wet the hydrophobic PDMS micropattern. In
total, 33 movies of A. castellanii migrating
on the samples were collected using a 60× oil immersion objective
(UPlanSApo 60× Oil, Olympus, Japan) on an inverted microscope
(IX-81, Olympus, Japan) and a digital camera (Hamamatsu C9300, Hamamatsu,
Japan) at 10 fps.