Fluorescent reporters image data: Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries
Fluorescent reporters raw microscopy data related to Cardona et al., Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries. This dataset includes fluorescent reporters: - GFP:CAR-1 - SPN-4:GFP Reference Cardona et al., Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries, Cell (2023), https://doi.org/10.1016/j.cell.2023.08.018
Steps to reproduce
Live C. elegans nematodes were anesthetized with 50 mM NaN3 in Phosphate Buffer Saline (PBS), and transferred between slides and coverslips to 4% agarose pads, and imaged on a Zeiss LSM 880 microscope with a Plan-Apochromat 40x/1.4 Oil DIC M27 objective and AiryScan SR module. Imaging was conducted in the first 15 min of anesthesia. To avoid signal interference from the intestine, images of GFP-tagged proteins were acquired from the gonad closer to the objective. 3D image data was obtained using Zeiss ZEN software and a 488 nm laser (BP 420-480 + BP 495-550). Z-stacks were acquired with a step-size of 0.5 µm (2.0 optical zoom, 0.66 μs pixel dwell, master gain of 750, digital offset of 1,79 µm pinhole) and processed using the in-built Zeiss Airyscan Processing algorithm.