Confocal imaging was performed on the Cured samples by a Zeiss LSM710 using a Plan-Apochrombat 63X/1.40 and an alpha Plan-APO 100X/1.46 oil objective lens (Carl Zeiss). 3D SIM imaging was performed with a Zeiss LSM710 Elyra P1 using an alpha Plan-APO 100X/1.46 oil objective lens. A laser at 561 nm (200 mW) was used for Alexa 568 (Invitrogen). A z-stack was acquired from 42 optical sections with a 200 nm interval. Each section was imaged using five rotations with a 51 nm grating period. 3D SIM Images were rendered using Zen 2012 SP2 software.
Zhao Y., Wang H., Wiesehoefer C., Shah N.B., Reetz E., Hwang J.Y., Huang X., Wang T.E., Lishko P.V., Davies K.M., Wennemuth G., Nicastro D, & Chung J.J. (2022). 3D structure and in situ arrangements of CatSper channel in the sperm flagellum. Nature Communications, 13, 3439.
+ Open protocol