4D electron microscopy of T cell activation

Yue Lu; Byung Kuk Yoo; Alphonsus H.C. Ng; Jungwoo Kim; Sinchul Yeom; Jau Tang; Milo M. Lin; Ahmed H. Zewail; James R. Heath

doi:10.1073/pnas.1914078116

4D electron microscopy of T cell activation

Yue Lu, Byung Kuk Yoo, Alphonsus H.C. Ng, Jungwoo Kim, Sinchul Yeom, Jau Tang, Milo M. Lin, Ahmed H. Zewail, James R. Heath

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

T cells can be controllably stimulated through antigen-specific or nonspecific protocols. Accompanying functional hallmarks of T cell activation can include cytoskeletal reorganization, cell size increase, and cytokine secretion. Photon-induced near-field electron microscopy (PINEM) is used to image and quantify evanescent electric fields at the surface of T cells as a function of various stimulation conditions. While PINEM signal strength scales with multiple of the biophysical changes associated with T cell functional activation, it mostly strongly correlates with antigen-engagement of the T cell receptors, even under conditions that do not lead to functional T cell activation. PINEM image analysis suggests that a stimulationinduced reorganization of T cell surface structure, especially over length scales of a few hundred nanometers, is the dominant contributor to these PINEM signal changes. These experiments reveal that PINEM can provide a sensitive label-free probe of nanoscale cellular surface structures.

Original language	English
Pages (from-to)	22014-22019
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	116
Issue number	44
DOIs	https://doi.org/10.1073/pnas.1914078116
State	Published - Oct 29 2019
Externally published	Yes

Funding

ACKNOWLEDGMENTS. We thank D. Baltimore for providing MART-1-specific Jurkat cells and K. A. Woodrow for providing the Zetasizer. Funding: This work was supported by the National Cancer Institute of the National Institutes of Health (U01CA217655). The authors thank the Parker Institute for Cancer Immunotherapy (PICI) for support. A.H.C.N. is supported by a Banting Postdoctoral Fellowship from the Government of Canada. M.M.L. acknowledges the Welch Foundation for support. We thank D. Baltimore for providing MART-1-specific Jurkat cells and K. A. Woodrow for providing the Zetasizer. Funding: This work was supported by the National Cancer Institute of the National Institutes of Health (U01CA217655). The authors thank the Parker Institute for Cancer Immunotherapy (PICI) for support. A.H.C.N. is supported by a Banting Postdoctoral Fellowship from the Government of Canada. M.M.L. acknowledges the Welch Foundation for support.

Keywords

Electron microscopy
Electron-photon coupling
PINEM
T cell activation
T cell receptor

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10.1073/pnas.1914078116

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title = "4D electron microscopy of T cell activation",

abstract = "T cells can be controllably stimulated through antigen-specific or nonspecific protocols. Accompanying functional hallmarks of T cell activation can include cytoskeletal reorganization, cell size increase, and cytokine secretion. Photon-induced near-field electron microscopy (PINEM) is used to image and quantify evanescent electric fields at the surface of T cells as a function of various stimulation conditions. While PINEM signal strength scales with multiple of the biophysical changes associated with T cell functional activation, it mostly strongly correlates with antigen-engagement of the T cell receptors, even under conditions that do not lead to functional T cell activation. PINEM image analysis suggests that a stimulationinduced reorganization of T cell surface structure, especially over length scales of a few hundred nanometers, is the dominant contributor to these PINEM signal changes. These experiments reveal that PINEM can provide a sensitive label-free probe of nanoscale cellular surface structures.",

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AU - Yoo, Byung Kuk

AU - Ng, Alphonsus H.C.

AU - Kim, Jungwoo

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AU - Tang, Jau

AU - Lin, Milo M.

AU - Zewail, Ahmed H.

AU - Heath, James R.

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AB - T cells can be controllably stimulated through antigen-specific or nonspecific protocols. Accompanying functional hallmarks of T cell activation can include cytoskeletal reorganization, cell size increase, and cytokine secretion. Photon-induced near-field electron microscopy (PINEM) is used to image and quantify evanescent electric fields at the surface of T cells as a function of various stimulation conditions. While PINEM signal strength scales with multiple of the biophysical changes associated with T cell functional activation, it mostly strongly correlates with antigen-engagement of the T cell receptors, even under conditions that do not lead to functional T cell activation. PINEM image analysis suggests that a stimulationinduced reorganization of T cell surface structure, especially over length scales of a few hundred nanometers, is the dominant contributor to these PINEM signal changes. These experiments reveal that PINEM can provide a sensitive label-free probe of nanoscale cellular surface structures.

KW - Electron microscopy

KW - Electron-photon coupling

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KW - T cell receptor

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4D electron microscopy of T cell activation

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