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Intravital Confocal Microscopy of Dermal Innate Immune Responses to Flea-Transmitted Yersinia pestis

  • Jeffrey G. ShannonEmail author
  • B. Joseph Hinnebusch
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2010)

Abstract

The technique known as intravital microscopy (IVM), when used in conjunction with transgenic mice expressing fluorescent proteins in various cell populations, is a powerful tool with the potential to provide new insights into host-pathogen interactions in infectious disease pathogenesis in vivo. Yersinia pestis, the causative agent of plague, is typically deposited in a host’s skin during feeding of an infected flea. IVM has been used to characterize the innate immune response to Y. pestis in the skin and identify differences between the responses to needle-inoculated and flea-transmitted bacteria that would have been difficult, if not impossible, to detect by other means. Here we describe techniques used to image the neutrophil response to flea-transmitted Y. pestis in the dermis of live mice using conventional confocal microscopy.

Key words

Intravital microscopy Confocal microscopy Skin Yersinia pestis Xenopsylla cheopis Flea 

Notes

Acknowledgements

We thank Clayton Jarrett, Chris Bosio, Dave Bland, Dustin Van Hofwegen, Adelaide Miarinjara, and Ashley Schwarzer for their careful review of the manuscript, David Dorward and Vinod Nair for assistance with confocal microscopy, and Anita Mora for assistance with graphic arts. This research was supported by the Intramural Research Program of the National Institutes of Health, National Institute of Allergy and Infectious Diseases.

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Copyright information

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

Authors and Affiliations

  1. 1.Laboratory of Bacteriology, Rocky Mountain LaboratoriesNIAID, NIHHamiltonUSA

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