Microscopy Images for: UTag, a cysteine-free thermostable tagging system for tracking single mRNA translation live

This dataset contains live-cell confocal microscopy images (OME-TIFF format) accompanying the manuscript "UTag, a cysteine-free thermostable tagging system for tracking single mRNA translation live." The repository includes 187 maximum-intensity z-projected time-lapse image files (2.6 GB (compressed) or ~13 GB (uncompressed)) organized into two subdirectories corresponding to distinct experimental analyses: Fig_7_ACF/ (Figures 7 F–I; 60 files, ~11 GB): Extended time-lapse recordings used for temporal autocorrelation analysis of translation spot fluorescence fluctuations. Fig_7_Harringtonine/ (Figures 7 J–M; 127 files, ~2 GB): Time-lapse recordings of cells treated with harringtonine. Each subdirectory is organized by tagging system: UTag, UTag_CF (cysteine-free variant), SunTag, and AlfaTag. And further subdivided by experimental date and replicate. Individual files are named by acquisition date, plasmid construct, and series number (e.g., 20250221_pNZ208_377_CAAX_Series001_maxZ.ome.tif). All live-cell images were acquired from U-2 OS cells using a Leica Stellaris 5 confocal microscope equipped with a 63× oil-immersion objective. During imaging, cells were maintained in a stage-top incubator at 37 °C, 5% CO₂, and 51% relative humidity. Images were collected with a 512 × 512-pixel field of view at 16-bit depth, yielding a pixel size of 129.89 nm. For z-stack acquisition, a step size of 0.3 µm was used. The files provided here are maximum-intensity z-projections of the acquired stacks. All image acquisition was performed using Leica Application Suite X (LAS X, version 4.5.0.25531). Data was processed using the code provided in this repository: https://github.com/ningzhaoAnschutz/utag_paper