Csu x1 spinning disk

Embryos were collected at 25 degrees overnight. They were then dechorionated with 50% bleach and washed with water. Embryos at the right stage were chosen under the fluorescent lamp and mounted depending on the experiment to be performed. For confocal imaging, the embryos were mounted on a glass-bottomed culture dish (Mattek), covered with 1% agarose and immersed in 1% PBS. For stretching experiments, the embryos were mounted on a glass-bottomed culture dish (Mattek) that had been coated with heptane glue, and immersed in 1% PBS. The imaging was carried using a Yokogawa CSU-X1 spinning disk on an inverted Olympus 1X81 microscope, with lens U plan S Apo 60x 1.45 Oil.

PhoCl activation was performed on an inverted Olympus IX71 microscope equipped with a Yokogawa CSU-X1 spinning disk; a ×60/1.42 numerical aperture Olympus oil objective was used, together with lasers at 491 nm (100 mW, Cobolt) and 561 nm (100 mW; Cobolt). A quad-edge dichroic beam splitter (446/523/600/677 nm; Semrock) was used to separate fluorescence emission from excitation light, and final images were taken with an Orca Flash 4.0 sCMOS camera (Hamamatsu). Images were acquired with MetaMorph (Molecular Devices).
All TIRF single-molecule-tracking experiments were performed on a custom-built microscope^{29 (link)}. An Olympus TIRF objective (×60/1.49 numerical aperture) was used with a 473-nm laser (100 mW; Laserglow Technologies) and a 643-nm laser (150 mW; Toptica Photonics). A quad-edge dichroic beam splitter (405/488/561/635 nm; Semrock) separated fluorescence emission from excitation light. Emission light was further filtered by a quad-band bandpass filter (446/523/600/677 nm; Semrock) and focused by a 500-mm tube lens onto the chip of a back-illuminated, electron-multiplying, charge-coupled device camera (Evolve, Photometrics) that was water-cooled to −85 °C. Images were acquired with MicroManager 2.0 (ref. ^{30 (link)}).