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. Author manuscript; available in PMC: 2021 Mar 1.
Published in final edited form as: Curr Protoc Cell Biol. 2020 Mar;86(1):e102. doi: 10.1002/cpcb.102

Table 2:

Example protocols for optogenetic experiments

Goal: To confirm presence and functionality of optogenetic probes: Monolayer (imaging mCherry Larg and Cell Mask Deep Red) : Single Cell (imaging mApple-Myosin and Alexa 647 Beads for Traction Force Microscopy):
Journal: Acquire mCherry (GEF) Timelapse Loop (30 second intervals for 10 min)

  • Acquire mCherry

  • Acquire Deep Red

 Timelapse Loop (20 second intervals for 15 min)

  • Acquire mApple

  • Acquire Alexa647
Acquire GFP (Stargazin) Timelapse Loop (30 second intervals for 5 min)

  • Mosaic Illumination in ROI 405 nm

  • Acquire mCherry

  • Acquire Deep Red

 Timelapse Loop (20 second intervals for 15 min)

  • Mosaic Illumination in ROI with 405 nm

  • Acquire mApple

  • Acquire Alexa647
Acquire mCherry (GEF) Timelapse Loop (30 second intervals for 15 min)

  • Acquire mCherry

  • Acquire Deep Red

 Timelapse Loop (20 second intervals for 15 min)

  • Acquire mApple

  • Acquire Alexa647
Confirm that the GEF moves from the cytosol to the membrane Remove cells from gels with 0.05% SDS
Acquire Alexa 647 (Relaxed gel image for TFM)