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Title:Centrosome polarization delivers secretory granules to the immunological synapse.
Authors:Stinchcombe JC, Majorovits E, Bossi G, Fuller S, Griffiths GM
Sample:Immunological synapse between a cytotoxic T lymphocyte and a target cell
Aggregation state:Individual structure (50 angstroms resolution)
Red flagLatest update:2011-05-26
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bullet_white.gif  Summary
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bullet.png Experimental details
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Sample
Sample name: Immunological synapse between a cytotoxic T lymphocyte and a target cell
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1cellular-componenthuman CD8 CTL clone, PK-10falsehomo sapiens0; human CD8 CTL clone derived from healthy donor
2cellular-componentP815 mouse target cell0falsehomo sapiens0
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
mg/mLRPMI mediumCTLs were labelled overnight with 1-2mg/ml HRP to load the lytic granules. After 30 min. conjugation to P815 target cells with anti-CD3 antibodies the sample was fixed and processed for DAB-cytochemistry and post-fixed with reduced osmium. The serial section was stained with lead citrate for 8 min.
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% K ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F30300 kVFLOOD BEAMBRIGHT FIELD2.2 mm4800 nm5200 nm56006200FIELD EMISSION GUNGATAN 2kx2k CCD camera mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
flip-flop holderOTHER-68.0°68.0°GIF0-20 eV K K K mrad e/Å2double tilt series2005-11-08
Processing
Software:IMOD
Resolution by author:50 Å
Resolution method:estimation - resolution is not isotropic!
Processing details:dual-axis tomograms were matched and merged to give a final map with reduced missing "cone"
Unit cell:
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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