Materials
| Name | Company | Catalog Number | Comments |
| #1.5 cover slips | VWR | 48393-172 | |
| BD Cytofix/Cytoperm | BD Biosciences | 554722 | |
| Bovine serum albumin | Sigma | A2153 | |
| Cotton tipped applicator | Fisher Scientific | S450941 | |
| Falcon centrifuge tubes (50 ml) | VWR | 352070 | |
| Fetal calf serum (FCS) (500 ml) | Atlantic Biologicals | S11050 | |
| Goat anti-rabbit Pacific Orange | Life Technologies | P31584 | |
| Laboratory tissue wipers | VWR | 82003-820 | |
| Nail polish | VWR | 100491-940 | |
| NK-92 cells | ATCC | CRL-2407 | |
| Phalloidin Alexa Fluor 488 | Life Technologies | A12379 | |
| Phosphate buffered saline | Life Technologies | 14190250 | |
| Prolong anti-fade reagent | Life Technologies | P7481 | |
| Purified anti-CD18 | Biolegend | 301202 | |
| Purified anti-NKp30 | Biolegend | 325202 | |
| Purified anti-perforin | Biolegend | 308102 | |
| RPMI 1640 medium (500 ml) | Life Technologies | 11875-093 | |
| Saponin from Quillaja bark | Sigma | S4521 | |
| Super PAP pen | Life Technologies | 008899 | |
| Triton X-100 | Electron Microscopy Sciences | 22142 | |
| Huygens deconvolution software | SVI | Contact company | |
| Leica SP8 TCS STED microscope | Leica Microsystems | contact company |
References
- Stinchcombe, J. C., Bossi, G., Booth, S., Griffiths, G. M. The immunological synapse of CTL contains a secretory domain and membrane bridges. Immunity. 15 (5), 751-761 (2001).
- Grakoui, A., Bromley, S. K., Sumen, C., et al. The immunological synapse: a molecular machine controlling T cell activation. Science. 285 (5425), 221-227 (1999).
- Monks, C. R., Freiberg, B. A., Kupfer, H., Sciaky, N., Kupfer, A. Three-dimensional segregation of supramolecular activation clusters in T cells. Nature. 395 (6697), 82-86 (1998).
- Orange, J. S., Harris, K. E., Andzelm, M. M., Valter, M. M., Geha, R. S., Strominger, J. L. The mature activating natural killer cell immunologic synapse is formed in distinct stages. Proc. Natl. Acad. Sci. U.S.A. 100 (24), 14151-14156 (2003).
- Treanor, B., Lanigan, P. M., Kumar, S., et al. Microclusters of inhibitory killer immunoglobulin-like receptor signaling at natural killer cell immunological synapses. J. Cell Biol. 174 (1), 153-161 (2006).
- Abeyweera, T. P., Merino, E., Huse, M. Inhibitory signaling blocks activating receptor clustering and induces cytoskeletal retraction in natural killer cells. J. Cell Biol. 192 (4), 675-690 (2011).
- Beemiller, P., Jacobelli, J., Krummel, M. F. Integration of the movement of signaling microclusters with cellular motility in immunological synapses. Nat. Immunol. 13 (8), 787-795 (2012).
- Campi, G., Varma, R., Dustin, M. L. Actin and agonist MHC-peptide complex-dependent T cell receptor microclusters as scaffolds for signaling. J. Exp. Med. 202 (8), 1031-1036 (2005).
- Mossman, K. D., Campi, G., Groves, J. T., Dustin, M. L. Altered TCR signaling from geometrically repatterned immunological synapses. Science. 310 (5751), 1191-1193 (2005).
- Varma, R., Campi, G., Yokosuka, T., Saito, T., Dustin, M. L. T cell receptor-proximal signals are sustained in peripheral microclusters and terminated in the central supramolecular activation cluster. Immunity. 25 (1), 117-127 (2006).
- Yokosuka, T., Sakata-Sogawa, K., Kobayashi, W., et al. Newly generated T cell receptor microclusters initiate and sustain T cell activation by recruitment of Zap70 and SLP-76. Nat. Immunol. 6 (12), 1253-1262 (2005).
- Toomre, D., Bewersdorf, J. A new wave of cellular imaging. Annu. Rev. Cell. Dev. Biol. 26, 285-314 (2010).
- Schermelleh, L., Heintzmann, R., Leonhardt, H. A guide to super-resolution fluorescence microscopy. J. Cell Biol. 190 (2), 165-175 (2010).
- Huang, B., Bates, M., Zhuang, X. Super-resolution fluorescence microscopy. Annu. Rev. Biochem. 78, 993-1016 (2009).
- Mace, E. M., Orange, J. S. New views of the human NK cell immunological synapse: recent advances enabled by super- and high-resolution imaging techniques. Front. Immunol. 3, 421 (2012).
- Hell, S. W., Wichmann, J. Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19 (11), 780-782 (1994).
- Klar, T. A., Hell, S. W. Subdiffraction resolution in far-field fluorescence microscopy. Opt. Lett. 24 (14), 954-956 (1999).
- Klar, T. A., Jakobs, S., Dyba, M., Egner, A., Hell, S. W. Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission. Proc. Natl. Acad. Sci. U.S.A. 97 (15), 8206-8210 (2000).
- Rankin, B. R., Moneron, G., Wurm, C. A., et al. Nanoscopy in a living multicellular organism expressing GFP. Biophys. J. 100 (12), 63-65 (2011).
- Willig, K. I., Kellner, R. R., Medda, R., Hein, B., Jakobs, S., Hell, S. W. Nanoscale resolution in GFP-based microscopy. Nat. Methods. 3 (9), 721-723 (2006).
- Rak, G. D., Mace, E. M., Banerjee, P. P., Svitkina, T., Orange, J. S. Natural killer cell lytic granule secretion occurs through a pervasive actin network at the immune synapse. PLoS Biol. 9 (9), (2011).
- Mace, E. M., Orange, J. S. Dual channel STED nanoscopy of lytic granules on actin filaments in natural killer cells. Commun. Integr. Biol. 5 (2), 184-186 (2012).
- Singh, H., Lu, R., Rodriguez, P. F., et al. Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy. Mitochondrion. 12 (2), 230-236 (2012).
- Tonnesen, J., Nagerl UV, Superresolution imaging for neuroscience. Exp. Neurol. 242, 33-40 (2013).
- Kempf, C., Staudt, T., Bingen, P., et al. Tissue multicolor STED nanoscopy of presynaptic proteins in the calyx of held. PLoS One. 8 (4), (2013).
- Jans, D. C., Wurm, C. A., Riedel, D., et al. STED super-resolution microscopy reveals an array of MINOS clusters along human mitochondria. Proc. Natl. Acad. Sci. U.S.A. 110 (22), 8936-8941 (2013).
- Gustafsson, M. G. Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J. Microsc. 198 (2), 82-87 (2000).
- Gustafsson, M. G., Shao, L., Carlton, P. M., et al. Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination). Biophys. J. 94 (12), 4957-4970 (2008).
- Schermelleh, L., Carlton, P. M., Haase, S., et al. Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy. Science. 320 (5881), 1332-1336 (2008).
- Betzig, E., Patterson, G. H., Sougrat, R., et al. Imaging intracellular fluorescent proteins at nanometer resolution. Science. 13 (5793), 1642-1645 (2006).
- Hess, S. T., Girirajan, T. P., Mason, M. D. Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys. J. 91 (11), 4258-4272 (2006).
- Rust, M. J., Bates, M., Zhuang, X. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy. Nat. Methods. 3 (10), 793-795 (2006).
- Cdc42-interacting protein-4 functionally links actin and microtubule networks at the cytolytic NK cell immunological. J. Exp. Med. Banerjee, P. P., Pandey, R., Zheng, R., Suhoski,, Monaco-Shawver, L., Orange, J. S. 204 (10), 2305-2320 (2007).
- Vicidomini, G., Moneron, G., Han, K. Y., et al. Sharper low-power STED nanoscopy by time gating. Nat. Methods. 8 (7), 571-573 (2011).
