Main description:

This new collection features the most up-to-date essential protocols that are currently being used to study the immune synapse. Beginning with methods for making biophysical measurements, the volume continues by covering the cell biology of synapses, methods for advanced substrate engineering, mechanobiology topics, new technologies to describe and manipulate synaptic components, as well as methods related to sites of action and immunotherapy. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and fully updated, The Immune Synapse: Methods and Protocols, Second Edition serves as an ideal practical guide for researchers working in this dynamic field.
Chapters 5, 11, 18, 27, 30, and 32 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Contents:

1. Measuring the Co-Localization and Dynamics of Mobile Proteins in Live Cells Undergoing Signaling Responses

Sarah A. Shelby, Thomas R. Shaw, and Sarah L. Veatch

2. Fluorescence-Based Measurements of Two-Dimensional Affinity in Membrane Interfaces

Tommy Dam, Manto Chouliara, and Peter Joensson

3. Surfaces for Study of Receptor Dynamics on T Cells

James McColl and David Klenerman

4. Molecular Dynamics Simulations of Immune Receptors and Ligands

Prithvi R. Pandey, Bartosz Rozycki, and Thomas R. Weikl

5. Investigating Diffusion Dynamics and Interactions with Scanning Fluorescence Correlation Spectroscopy (sFCS)

Alexander M. Morch and Falk Schneider

6. Combined FRET-FLIM and NAD(P)H FLIM to Analyze B Cell Receptor Signaling Induced Metabolic Activity of Germinal Center B Cells In Vivo

Carolin Ulbricht, Ruth Leben, Yu Cao, Raluca A. Niesner, and Anja E. Hauser

7. Measurement of Molecular Height Using Cell Surface Optical Profilometry (CSOP)

Sungmin Son and Daniel A. Fletcher

8. Observing Membrane and Cell Adhesion via Reflection Interference Contrast Microscopy (RICM)

Ahmed Abdelrahman, Ana-Suncana Smith, and Kheya Sengupta

9. En-Face-Imaging of T Cell-Dendritic Cell Immunological Synapses

Alexander Leithner, Jack Merrin, and Michael Sixt

10. High and Super Resolution Imaging of Cell-Cell Interfaces

Julia Sajman and Eilon Sherman

11. Separation of Single Core and Multicore Lytic Granules by Subcellular Fractionation and Immuno-Isolation

Claudia Schirra, Nadia Alawar, Ute Becherer, and Hsin-Fang Chang

12. Microvillar Cartography: A Super-Resolution Single-Molecule Imaging Method to Map the Positions of Membrane Proteins with Respect to Cellular Surface Topography

Shirsendu Ghosh, Andres Alcover, and Gilad Haran

13. T Cell Immunological Synaptosomes: Definition and Isolation

Hye-Ran Kim, Jeong-Su Park, Na-Young Kim, and Chang-Duk Jun

14. Dynamics of Immune Cell Microvilli

En Cai

15. Visualization of Myddosome Assembly in Live Cells

Fakun Cao and Marcus J. Taylor

16. Detection of Telomere Transfer at Immunological Synapse

Alessio Lanna and Clara D'Ambra

17. Bottom-Up Assembly of Bio-Inspired, Fully Synthetic Extracellular Vesicles

Meline Macher, Ilia Platzman, and Joachim P. Spatz

18. A DNA Origami-Based Biointerface to Interrogate the Spatial Requirements for Sensitized T-Cell Antigen Recognition

Joschka Hellmeier, Rene Platzer, Johannes B. Huppa, and Eva Sevcsik

19. Leveraging DNA Origami to Study Phagocytosis

Wyatt D. Miller, Nadja Kern, Shawn M. Douglas, and Meghan A. Morrissey

20. Fabrication of Nanoscale Arrays to Study the Effect of Ligand Arrangement on Inhibitory Signaling in NK Cells

Guillaume Le Saux, Esti Toledo-Ashkenazi, and Mark Schvartzman

21. Measurement of Forces for Trans-Endocytosis at Dorsal and Ventral Sides of the Cell

Seungman Park and Yun Chen

22. Functionalized Lipid Droplets and Microfluidics Approach to Study Immune Cell Polarity In Vitro

Judith Pineau, Lea Pinon, Jacques Fattaccioli, and Paolo Pierobon

23. Quantifying Immune Cell Force Generation Using Traction Force Microscopy

Marcel Issler, Huw Colin-York, and Marco Fritzsche

24. Characterizing Biophysical Parameters of Single TCR-pMHC Interactions Using Optical Tweezers

Hannah M. Stephens, Evan Kirkpatrick, Robert J. Mallis, Ellis L. Reinherz, and Matthew J. Lang

25. Isolation of the B Cell Immune Synapse for Proteomic Analysis

Diogo M. Cunha, Sara Hernandez-Perez, and Pieta K. Mattila

26. Analyzing Single Cell Secretions by 'Shadow Imaging'

Ashley R. Ambrose, Khodor S. Hazime, and Daniel M. Davis

27. Exploiting the RUSH System to Study Lytic Granule Biogenesis in Cytotoxic T Lymphocytes

Nagaja Capitani, Chiara Cassioli, Keerthana Ravichandran, and Cosima T. Baldari

28. Interactions of Tissue Resident T Cells

Rut Mora-Buch, Hasan Akbaba, and Shannon K. Bromley

29. Live Imaging of CAR T Cell Ca2+ Signals in Tumor Slices Using Confocal Microscopy

David Espie, Sarah Barrin, Irena Rajnpreht, Lene Vimeux, and Emmanuel Donnadieu

30. Measuring CTL Lytic Granule Secretion and Target Cell Membrane Repair by Fluorescent Lipophilic Dye Uptake at the Lytic Synapse

Sabina Muller, Liza Filali, Marie-Pierre Puissegur, and Salvatore Valitutti

31. In Vitro Generation of Human Tolerogenic Monocyte-Derived Dendritic Cells

Catharien M.U. Hilkens, Julie Diboll, Fiona Cooke, and Amy E. Anderson

32. Methods of Machine Learning-Based Chimeric Antigen Receptor Immunological Synapse Quality Quantification

Julian Gan, Jong Hyun Cho, Ryan Lee, Alireza Naghizadeh, Ling Yue Poon, Ethan Wang, Zachary Hui, and Dongfang Liu

33. Imaging CAR-T Synapse as a Quality Control for CAR Engineering

Qian Xiao and Xiaolei Su