June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Depot Indocyanine Green Dye for in vivo visualization of infiltrating leukocytes
Author Affiliations & Notes
  • Dawn A Sim
    NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom
    UCL Institute of Ophthalmology, London, United Kingdom
  • Colin J Chu
    UCL Institute of Ophthalmology, London, United Kingdom
  • Michael B Powner
    UCL Institute of Ophthalmology, London, United Kingdom
  • Senthil Selvam
    UCL Institute of Ophthalmology, London, United Kingdom
  • Adnan Tufail
    NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom
    UCL Institute of Ophthalmology, London, United Kingdom
  • Catherine A Egan
    NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom
  • James W B Bainbridge
    NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom
    UCL Institute of Ophthalmology, London, United Kingdom
  • Richard W J Lee
    NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom
    Academic unit of Ophthalmology, School of Clinical Sciences, University of Bristol, Bristol, United Kingdom
  • Andrew D Dick
    NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom
    Academic unit of Ophthalmology, School of Clinical Sciences, University of Bristol, Bristol, United Kingdom
  • Marcus Fruttiger
    UCL Institute of Ophthalmology, London, United Kingdom
  • Footnotes
    Commercial Relationships Dawn Sim, Allergan (F), UCL Business (P); Colin Chu, None; Michael Powner, None; Senthil Selvam, None; Adnan Tufail, UCL Business (P); Catherine Egan, None; James Bainbridge, None; Richard Lee, None; Andrew Dick, None; Marcus Fruttiger, UCL Business (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4117. doi:
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      Dawn A Sim, Colin J Chu, Michael B Powner, Senthil Selvam, Adnan Tufail, Catherine A Egan, James W B Bainbridge, Richard W J Lee, Andrew D Dick, Marcus Fruttiger; Depot Indocyanine Green Dye for in vivo visualization of infiltrating leukocytes. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4117.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

To assess a novel method to label and image myeloid cells infiltrating the mouse retina and choroid in vivo, using a single depot injection of Indocyanine green dye (ICG).

 
Methods
 

We assess the use of depot ICG for the purpose of in vivo cell labelling on three different ocular models of inflammation and angiogenesis - endotoxin-induced uveitis (EIU), experimental autoimmune uveoretinitis (EAU) and laser-induced choroidal neovascularisation (CNV) model. A near-infrared scanning ophthalmoscope was used for in vivo imaging of the eye and flow cytometry was used on blood and spleen to assess the number and phenotype of labelled cells. ICG was administered 72 hours prior to the induction of inflammation to ensure clearance from systemic circulation.

 
Results
 

We found that in vivo intravenous administration failed to label any leukocytes, whereas depot injection, either intraperitoneal or subcutaneous, was successful in labelling CD11b+ myeloid cells. Progression of inflammation could be traced over a period of 14 days following a single depot injection of ICG. Additionally, flow cytometric analysis revealed that the predominant population of cells stained by ICG are circulating and splenic reservoir CD11b+ myeloid cells.

 
Conclusions
 

The translation of this approach into clinical practice would enable visualisation of immune cells in situ. This will not only provide a greater understanding of pathogenesis, monitoring and assessment of therapy in many human ocular diseases but also opens the ability to image immunity live for neurodegenerative disorders, cardiovascular disease and systemic immune mediated disorders.  

 
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