smFISH image data (part 3): Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries
Description
smFISH raw microscopy data related to Cardona et al., Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries. This dataset includes: 1. One-color smFISH image data: - Channel 1: Cy3 labeled mRNA target spn-4, glp-1, puf-5, tbb-2, or pccb-1. - If present, Channel 2: GFP labeled macro-condensate marker CAR-1. 2. Two-color smFISH image data: - Channel 1: Cy5 labeled mRNA target. - Channel 2: Cy3 labeled mRNA target. - If present, Channel 3: GFP labeled macro-condensate marker CAR-1. 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
Files
Steps to reproduce
smFISH images were acquired on a Zeiss LSM 880 microscope with a Plan-Apochromat 63x/1.4 Oil DIC M27 objective. Images were processed with the AiryScan super-resolution (SR) module, using Zeiss ZEN software. A 561 nm laser (BP 570-620 + LP 645 + SBS SP 615) was used for mRNA detection, and a 488 nm laser (BP 420-480 + BP 495-550) for GFP:CAR-1 signal to assess mRNA localization into P-bodies that are GFP:CAR-1 labelled. For AiryScan acquisitions, images were taken with a z-spacing of 0.185 μm, 2.0 optical zoom, 0.66 μs pixel dwell, average of 4, master gain of 750, digital offset of 1, and 143 µm pinhole. For Fast AiryScan aquisitions, z-stacks were obtained with a step-size of 0.250 μm, 1.8 optical zoom, 0.52 μs pixel dwell, average of 4, master gain of 750, digital offset of 1, and 384 µm pinhole. Multiphase 3D smFISH images were processed using the in-built Zeiss Airyscan Processing algorithm.