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Three-Dimensional Structured Illumination Microscopy (3D-SIM) to Dissect Signaling Cross-Talks in Motile T-Cells

  • Seow Theng OngEmail author
  • Graham D. Wright
  • Navin Kumar Verma
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

Abstract

Visualization of signal transduction events in T-cells has always been a challenge due to their miniscule size. Recent advancement in super-resolution microscopy techniques presents many new opportunities to navigate the spatial and temporal signaling cross-talks in motile T-cells. Here, we provide technical details, optimal conditions, and critical practical considerations that need to be taken into account during cell handling, sample preparation, and image acquisition of motile T-cells for performing three-dimensional structured illumination microscopy (3D-SIM).

Key words

Super-resolution microscopy 3D-SIM Immunostaining 

Notes

Acknowledgments

This work was supported in part by grants from Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant and the Ministry of Education Singapore under its Singapore Ministry of Education Academic Research Fund (AcRF) Tier 2 Grant (MOE2017-T2-2-004) to N.K.V. 3D-SIM platform (DeltaVision OMX v4 Blaze microscope) and Institute of Medical Biology (IMB) Microscopy Unit, now renamed to the A*STAR Microscopy Platform within the Skin Research Institute of Singapore (SRIS), was funded by A*STAR, Singapore.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Seow Theng Ong
    • 1
    Email author
  • Graham D. Wright
    • 2
  • Navin Kumar Verma
    • 3
  1. 1.Lymphocyte Signalling Research Laboratory, Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore
  2. 2.A*STAR Microscopy PlatformSkin Research Institute of SingaporeSingaporeSingapore
  3. 3.Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore

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