Datasets for "Single-molecule and super-resolved imaging deciphers membrane behaviour of onco-immunogenic CCR5"

<strong>Flow cytometry</strong> Modality / instrument: <em>Flow cytometer</em> <em>(CytoFLEX LX, Beckman Coulter)</em> File format:<em> FCS + XIT (CytExpert, Beckman Coulter).</em> Samples and acquisitions: Fluorescent fusions in live Chinese Hamster ovary (CHO) cells. <em>File</em> <em>Cell line</em> <em>Runs</em> <em>Cells counted</em> CONTROL.fcs CHO wild-type 1 7000 GFP-CCR5.fcs CHO-GFP-CCR5 1 7000 Exp_20220916_1_GFP.xit N/A - metadata Approx. size 6 MB <strong>PaTCH microscopy images</strong> Imaging modality / instrument: <em>Brightfield</em> + <em>PaTCH fluorescence microscopy</em> Image format:<em> OME TIFF (16 bit) + MicroManager metadata files</em> Microscope settings: <em>488 nm triggered excitation; split red/green detection, cropped to green (GFP) channel only; 10 ms/frame laser exposure; 13.5 ms/frame-to-frame; 53 nm/px. Photometrics Prime95b CMOS.</em> Samples and acquisitions: Fluorescent fusions of GFP-CCR5 receptor in live CHO cells imaged with and without 100 nM CCL5 ligand. Each subfolder corresponds to a field of view and contains one brightfield and one PaTCH acquisition of the same cell. Folder Condition Fields of view AC6 CONTROL sc CCL5- 11 AC6 CCL5 sc CCL5+ (100 nM) 10 Approx. size before compression: 14 GB <strong>Structured illumination microscopy - volumetric stacks</strong> Imaging modality / instrument: <em>SIM fluorescence microscopy (custom setup at NPL based on Olympus IX71)</em> Image format:<em> OME TIFF (16 bit) with intrinsic metadata (voxel size)</em> Microscope settings: <em>638 nm excitation; 60x/1.3 NA; Flash 4.0, Hamamatsu Photonics. For additional details see the reference below (Hunter et al, bioRxiv).</em> Samples and acquisitions: Dylight 650-MC-5 labeled CCR5 receptor in fixed CHO-CCR5 cells, imaged with and without 100 nM CCL5 ligand. Each acquisition is of a unique field of view and contains one SIM reconstruction as an XYZ volumetric stack. ‘Basal membrane’ acquisitions consist of 5 slices at 200 nm z-intervals across the range of the basal membrane. ‘Whole cell' acquisitions are made up of 7 slices with 500 nm z-interval ranging from just below the basal membrane to just above the apical membrane. Folder Subfolder/condition Fields of view Basal membrane CCL5- 5 Basal membrane CCL5+ (100 nM) 6 Whole cells CCL5- 5 Whole cells CCL5+ (100 nM) 8 Approx. size before compression: 300 MB