May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Quantitative Characteristics of Fluorescein for Fluorescence Angiograms
Author Affiliations & Notes
  • A. Schwartz
    Center for Quantitative Cytometry, San Juan, Puerto Rico
  • C. Marques
    Association for Biomedical Research and Innovation on Light and Image, Coimbra, Portugal
  • G. Abreu
    Association for Biomedical Research and Innovation on Light and Image, Coimbra, Portugal
  • R. Benardes
    Association for Biomedical Research and Innovation on Light and Image, Coimbra, Portugal
  • L. Jolin
    Erie Scientific Co., Portsmouth, New Hampshire
  • E. Snow
    Erie Scientific Co., Portsmouth, New Hampshire
  • L. Wang
    National Institute of Standards and Technology, Gaithersburg, Maryland
  • A. Gaigalas
    National Institute of Standards and Technology, Gaithersburg, Maryland
  • W. Carter
    Pfizer, Inc., Groton, Connecticut
  • J. Cunha-Vaz
    Coimbra University Hospital, Coimbra, Portugal
  • Footnotes
    Commercial Relationships A. Schwartz, Consultant for Pfizer, Inc., C; Inventor designated on a patent application on presentation material, P; C. Marques, None; G. Abreu, None; R. Benardes, Inventor on patent application, P; L. Jolin, Company making standards, E; E. Snow, Company making standards, E; L. Wang, None; A. Gaigalas, None; W. Carter, Company funding the research, E; J. Cunha-Vaz, None.
  • Footnotes
    Support Pfizer, Inc. contract research
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 157. doi:https://doi.org/
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    • Get Citation

      A. Schwartz, C. Marques, G. Abreu, R. Benardes, L. Jolin, E. Snow, L. Wang, A. Gaigalas, W. Carter, J. Cunha-Vaz; Quantitative Characteristics of Fluorescein for Fluorescence Angiograms. Invest. Ophthalmol. Vis. Sci. 2007;48(13):157. doi: https://doi.org/.

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

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Abstract

Purpose:: Quantitation of image intensities from fluorescence angiograms could provide reliable information if they were independent of the instrument and the operator. The first step to obtaining such independent quantitative data is to ensure that the characteristics of fluorescein are consistent from different sources and traceable back to an accepted standard.

Methods:: The present study involved five different sources of fluorescein; one clinical injectable grade, three different "pure" sources from different companies, and the Standard Reference Material fluorescein (SRM 1932) from the National Institute of Standards and Technology. Moreover, sets of data were independently obtained from three separate laboratories; AIBILI in Coimbra, Portugal, Erie Scientific in Portsmouth, NH, and NIST in Gaithersburg, MD. Solutions of fluorescein (10-7 - 10-11 M) from each source were made in PBS at pH 7.4 and emission data were obtained with spectrofluorometers in each laboratory.

Results:: The intra-laboratory data from the five sources of fluorescein all described a single calibration line, as well as the same sigmoidal response as a function of pH from 5.0 to 9.0. Inter-laboratory data all described a single calibration line after being normalized, due to differences in the instruments, to the highest fluorescein concentration (800 ng/ml) that maintained linear response in the spectrofluorometer.

Conclusions:: Since all data from the three locations were equivalent, this indicates that fluorescein from different sources, including the injectable fluorescein, is traceable back to the NIST SRM 1932. To date, interpretation of fluorescent image intensities has been limited since it has been difficult to compare angiograms across laboratories and time. This multi-site, multi-source study is significant because it provides confidence that quantitative fluorescent measurements based on fluorescein standards made from different sources can provide comparable quantitative results.

Keywords: imaging methods (CT, FA, ICG, MRI, OCT, RTA, SLO, ultrasound) • imaging/image analysis: clinical • development 
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