Junction Tension Measurement via Myosin Sensor

To validate the SF9-3xGFP myosin sensor as a proxy for junction tension, we performed junction recoil measurements in stage 16 to stage 20 embryos expressing the SF9-3xGFP myosin II sensor. Laser ablations were performed using a 532nm pulse laser (>60 µJ pulses at 200Hz) at 20% power, with each ablation set for 5 ms. The ablation system was connected to a 3i spinning disk microscope with a Plan-Apochromat ×63 oil objective (N.A. = 1.4) mounted on an inverted Zeiss Axio Observer Z1 microscope (Marianas Imaging Workstation [3i—Intelligent Imaging Innovations]), equipped with a CSU-X1 spinning disk confocal head (Yokogawa) and an iXon Ultra 888 EM-CCD camera (Andor Technology). Vertex displacement was manually tracked using Fiji and the recoil velocities were calculated as previously described^{18 (link)}.

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Ventura G., Amiri A., Thiagarajan R., Tolonen M., Doostmohammadi A, & Sedzinski J. (2022). Multiciliated cells use filopodia to probe tissue mechanics during epithelial integration in vivo. Nature Communications, 13, 6423.

Publication 2022

CSU-X1 spinning disk confocal head iXon Ultra 888 EM-CCD camera

Embryos Innovations Laser ablations Microscope Myosin a Myosin ii Pulses

Corresponding Organization : University of Copenhagen

Other organizations : Novo Nordisk Foundation, Max Planck Institute for the Physics of Complex Systems

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Variable analysis

independent variables

Laser ablations were performed using a 532nm pulse laser (>60 µJ pulses at 200Hz) at 20% power, with each ablation set for 5 ms.

dependent variables

Vertex displacement
Recoil velocities

control variables

Stage 16 to stage 20 embryos expressing the SF9-3xGFP myosin II sensor
3i spinning disk microscope with a Plan-Apochromat ×63 oil objective (N.A. = 1.4) mounted on an inverted Zeiss Axio Observer Z1 microscope (Marianas Imaging Workstation [3i—Intelligent Imaging Innovations]), equipped with a CSU-X1 spinning disk confocal head (Yokogawa) and an iXon Ultra 888 EM-CCD camera (Andor Technology)

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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