In vivo fluorescent optical imaging of cytotoxic T lymphocyte migration using IRDye800CW near-infrared dye

Appl Opt. 2008 Nov 1;47(31):5944-52. doi: 10.1364/ao.47.005944.


We describe a method to measure in vivo migration of human T cells by using the near-infrared (NIR) dye IRDye800CW. Labeling of Epstein-Barr virus-specific T cells with IRDye800CW did not affect viability, proliferation, or T cell function. Following tail vein injection into mice bearing subcutaneous tumors, the NIR signal could be measured in vivo at the tumor site. Analysis of tumors revealed T cell infiltration and an increased NIR signal, confirming T cell migration. To test specific migration with IRDye800CW, tumors were modified to express CCL5 to measure site-specific migration. The NIR signal was increased at CCL5-secreting tumors compared with control tumors. Together, these data suggest that IRDye800CW may be used to study the trafficking of T cells in a small animal model and may have potential as a short-term reporter molecule for human immunotherapy studies.

MeSH terms

  • Animals
  • Cell Movement
  • Cell Proliferation
  • Cell Survival
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry / methods
  • Fluorescent Dyes / pharmacology*
  • Humans
  • Immunohistochemistry / methods
  • Mice
  • Optics and Photonics
  • Spectrometry, Fluorescence / methods
  • Spectroscopy, Near-Infrared / methods
  • T-Lymphocytes, Cytotoxic / cytology*


  • Fluorescent Dyes