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Modified Intravital Microscopy to Assess Vascular Health and T-Cell Motility

  • Geoffrey W. PayneEmail author
  • Kevin Mitchell
  • Stephanie L. Sellers
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

Abstract

The ability to study the microcirculation in real time is key to elucidating how the immune system and the associated microvasculature interact and influence one another within the lymph node (LN). Here, we present a method for near in-situ imaging of the inguinal LN. In particular, this method is ideal for the assessment of overall vascular health that influences immune functions and for the evaluation of T-cell motility. We focus on imaging of the microvasculature of the LN, paying particular attention to methods that ensure the study of healthy vessels, the ability to maintain imaging of viable vessels over a number of hours, quantification of vessel magnitude and vessel integrity. Modified intravital microscopy (M-IVM) of the LNs allows direct evaluation of microvascular functions as well as real-time imaging of the direct interface between immune cells, the LN, and the microcirculation. Importantly, M-IVM technique can be readily combined with many other vascular and immunological techniques such as fluorescent cell labeling and assessment of sticking and rolling time as descripted. Furthermore, it can be adapted to study vasculature of other than the inguinal LN. Overall, this chapter provides a dependable method for fundamental vascular immunological assessment of LNs that is decidedly useful in a diverse range of investigations.

Key words

Lymph node Intravital microscopy Microcirculation T-cell motility 

Notes

Acknowledgements

This work was supported by the Canadian Institutes of Health, Centre for Blood Research, Michael Smith Foundation and University of Northern British Columbia & University of British Columbia. SLS is supported by Michael Smith Foundation for Health Research/Centre for Heart Lung Innovation—Providence Health Care Research Institute—St. Paul’s Foundation Research Trainee Award.

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Copyright information

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

Authors and Affiliations

  • Geoffrey W. Payne
    • 1
    Email author
  • Kevin Mitchell
    • 1
  • Stephanie L. Sellers
    • 2
    • 3
  1. 1.Northern Medical ProgramUniversity of Northern British ColumbiaPrince GeorgeCanada
  2. 2.Centre for Heart Lung Innovation, St. Paul’s HospitalUniversity of British ColumbiaVancouverCanada
  3. 3.Department of Radiology, St. Paul’s HospitalUniversity of British ColumbiaVancouverCanada

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