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

Flow cytometry Modality / instrument: Flow cytometer (CytoFLEX LX, Beckman Coulter) File format: FCS + XIT (CytExpert, Beckman Coulter). Samples and acquisitions: Fluorescent fusions in live Chinese Hamster ovary (CHO) cells. File Cell line Runs Cells counted 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 PaTCH microscopy images Imaging modality / instrument: Brightfield + PaTCH fluorescence microscopy Image format: OME TIFF (16 bit) + MicroManager metadata files Microscope settings: 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. 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 Structured illumination microscopy - volumetric stacks Imaging modality / instrument: SIM fluorescence microscopy (custom setup at NPL based on Olympus IX71) Image format: OME TIFF (16 bit) with intrinsic metadata (voxel size) Microscope settings: 638 nm excitation; 60x/1.3 NA; Flash 4.0, Hamamatsu Photonics. For additional details see the reference below (Hunter et al, bioRxiv). 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
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