Computational protocol: Streptothricin acetyl transferase 2 (Sat2): A dominant selection marker for Caenorhabditis elegans genome editing

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Protocol publication

[…] The worm images on the plates were acquired using an SZX16 stereoscopic fluorescence microscope (Olympus) equipped with a CoolSNAP HQ cooled-CCD camera (Photometrics, Tucson, AZ, USA). Before image acquisition, the plates were cooled at −30°C for 1 min to prevent worm movements. For the fluorescence imaging of the sam-4::gfp integrated worms, we used an ORCA-R2 digital CCD camera (Hamamatsu Photonics, Hamamatsu, Japan) on an IX71 microscope (Olympus) with an UPlanSApo 60x/ NA1.3 silicone oil objective lens (Olympus) and a CSU-X1 spinning disc confocal system (Yokogawa Electric Corporation, Tokyo, Japan). Worms were mounted onto 3% agar pads with 2.5mM levamisol in M9 buffer, as described previously []. MetaMorph software (Molecular Devices, Sunnyvale, CA, USA) was used to control both microscopes. Fluorescent images of the worms were taken with 40 Z-slices and 1 μm intervals. ImageJ (NIH, https://imagej.nih.gov/ij/index.html) was used to process the images acquired. […]

Pipeline specifications

Software tools MetaMorph, ImageJ
Application Microscopic phenotype analysis
Organisms Caenorhabditis elegans
Chemicals Hygromycin B, Streptothricins