Noninvasive measurement of pharmacokinetics by near-infrared fluorescence imaging in the eye of mice

J Biomed Opt. 2014 Jan;19(1):16022. doi: 10.1117/1.JBO.19.1.016022.


Purpose: For generating preclinical pharmacokinetics (PKs) of compounds, blood is drawn at different time points and levels are quantified by different analytical methods. In order to receive statistically meaningful data, 3 to 5 animals are used for each time point to get serum peak-level and half-life of the compound. Both characteristics are determined by data interpolation, which may influence the accuracy of these values. We provide a method that allows continuous monitoring of blood levels noninvasively by measuring the fluorescence intensity of labeled compounds in the eye and other body regions of anesthetized mice.

Procedures: The method evaluation was performed with four different fluorescent compounds: (i) indocyanine green, a nontargeting dye; (ii) OsteoSense750, a bone targeting agent; (iii) tumor targeting Trastuzumab-Alexa750; and (iv) its F(ab')2-alxea750 fragment. The latter was used for a direct comparison between fluorescence imaging and classical blood analysis using enzyme-linked immunosorbent assay (ELISA).

Results: We found an excellent correlation between blood levels measured by noninvasive eye imaging with the results generated by classical methods. A strong correlation between eye imaging and ELISA was demonstrated for the F(ab')2 fragment. Whole body imaging revealed a compound accumulation in the expected regions (e.g., liver, bone).

Conclusions: The combination of eye and whole body fluorescence imaging enables the simultaneous measurement of blood PKs and biodistribution of fluorescent-labeled compounds.

MeSH terms

  • Animals
  • Antibodies, Monoclonal, Humanized / chemistry
  • Calibration
  • Enzyme-Linked Immunosorbent Assay
  • Eye / pathology*
  • Female
  • Fluorescence
  • Immunoglobulin Fragments / chemistry
  • Indocyanine Green / chemistry
  • Kinetics
  • Mice
  • Mice, SCID
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Reproducibility of Results
  • Spectroscopy, Near-Infrared / methods*
  • Succinimides / chemistry
  • Trastuzumab


  • Alexa Fluor 750
  • Antibodies, Monoclonal, Humanized
  • Immunoglobulin Fragments
  • Succinimides
  • Indocyanine Green
  • Trastuzumab