Table 1. Imaging parameters for the different experiments described.

Experiment	Objective	Fluorochrome(Excitation/emission wavelengths)	Image spacing in z	Exposure time per image	Number of Image stacks	Total time	Volume(x, y, z)
Microspheres (Matrigel)	60×	DragonGreen (490/507–530 nm)	41 slices, 1 µm apart	16ms	60	64 s	225.3 µm × 84.5 µm×40 µm
		tdTomato parasites (550/579–608 nm)
Fibrin vs Matrigel and TgMIC2 KO directionality	20×	Hoechst 33342 (385/420–449 nm)	41 x 1 µm	16ms	120	80 s	665.6 µm × 249.6 µm×40 µm
Force Mapping, WT (Fibrin)	60×	tdTomato parasites (550/579–608 nm)	50x0.5 µm	16ms	60	96 s	225.3 µm × 84.5 µm×24.5 µm
		Fibrin (635/666–723 nm)
WT, TgMyoA KO, TgMIC2 KO; Brightfield (Matrigel, fibrin)	60×	N/A: Brightfield	21 x 1 µm	40ms	360	302 s	225.3 µm × 225.3 µm×20 µm
Nuclear size vs constriction (Matrigel)	20×	Hoechst 33342 (385/420–449 nm)	41 x 1 µm	16ms	60	80 s	665.6 µm × 249.6 µm×40 µm
		Anti-SAG1 Alexa 548 (550/579–608 nm)
TgMyoA KO Force Map (Fibrin)	60×	Hoechst 33342 (385/420–449 nm)	50x0.5 µm	16ms	60	96 s	225.3 µm × 84.5 µm×24.5 µm
		Fibrin (635/666–723 nm)
TgMIC2 KO Force Map (Fibrin)	60×	YFP cytosol (490/507–530 nm)	50x0.5 µm	16ms	60	96 s	225.3 µm × 84.5 µm×24.5 µm
		Fibrin (635/666–723 nm)