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Real-time differential labeling of blood, interstitium, and lymphatic and single-field analysis of vasculature dynamics in vivo

Academic Article
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Overview

related to degree

  • Cahalan, Stuart Morgan, Ph.D. in Biology, Scripps Research 2004 - 2011

authors

  • Sarkisyan, G.
  • Cahalan, Stuart Morgan
  • Gonzalez-Cabrera, P. J.
  • Leaf, N. B.
  • Rosen, Hugh

publication date

  • May 2012

journal

  • American Journal of Physiology-Cell Physiology  Journal

abstract

  • Lymph nodes are highly organized structures specialized for efficient regulation of adaptive immunity. The blood and lymphatic systems within a lymph node play essential roles by providing functionally distinct environments for lymphocyte entry and egress, respectively. Direct imaging and measurement of vascular microenvironments by intravital multiphoton microscopy provide anatomical and mechanistic insights into the essential events of lymphocyte trafficking. Lymphocytes, blood endothelial cells, and lymphatic endothelial cells express sphingosine 1-phosphate receptor 1, a key G protein-coupled receptor regulating cellular egress and a modulator of endothelial permeability. Here we report the development of a differential vascular labeling (DVL) technique in which a single intravenous injection of a fluorescent dextran, in combination with fluorescent semiconductor quantum dot particles, differentially labels multiple blood and lymphatic compartments in a manner dependent on the size of the fluorescent particle used. Thus DVL allows measurement of endothelial integrity in multiple vascular compartments and the affects or pharmacological manipulation in vascular integrity. In addition, this technique allows for real-time observation of lymphocyte trafficking across physiological barriers differentiated by DVL. Last, single-field fluid movement dynamics can be derived, allowing for the simultaneous determination of fluid flow rates in diverse blood and lymphatic compartments.

subject areas

  • Animals
  • Computer Systems
  • Endothelial Cells
  • Extracellular Fluid
  • Lymph Nodes
  • Lymphatic Vessels
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Receptors, Lysosphingolipid
  • Regional Blood Flow
  • Staining and Labeling
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Research

keywords

  • fluorescence labeling
  • intravital two-photon excitation microscopy
  • lymph node
  • microfluidic dynamics
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Identity

International Standard Serial Number (ISSN)

  • 0363-6143

Digital Object Identifier (DOI)

  • 10.1152/ajpcell.00382.2011

PubMed ID

  • 22357735
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Additional Document Info

start page

  • C1460

end page

  • C1468

volume

  • 302

issue

  • 10

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