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Fluorescence Lifetime Imaging Ophthalmoscopy pp 23–34Cite as

Image Acquisition and Analysis

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Abstract

FLIO image acquisition follows a predefined standardized protocol. An autofluorescence intensity image and corresponding FLIO lifetime data are recorded in parallel. Becker & Hickl software is used for approximation of a decay curve for every single pixel. The mean fluorescence lifetime is calculated in both spectral channels. Extraction of fluorescence lifetime parameters is performed using FLIO reader or FLIMX software. Analysis software allow for data evaluation within a commonly used ETDRS grid as well as for specific regions of interest.

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References

  1. Dysli C, et al. Quantitative analysis of fluorescence lifetime measurements of the macula using the fluorescence lifetime imaging ophthalmoscope in healthy subjects. Invest Ophthalmol Vis Sci. 2014;55(4):2106–13.

    Article  Google Scholar 

  2. Dysli C, et al. Fluorescence lifetime imaging ophthalmoscopy. Prog Retin Eye Res. 2017;60:120–43.

    Article  Google Scholar 

  3. Matthias Klemm LS, Klee S, Link D, Peters S, Hammer M, Schweitzer D, Haueisen J. Bleaching effects and fluorescence lifetime imaging ophthalmoscopy. Biomed Opt Express. 2019;10(3):1446–61.

    Article  Google Scholar 

  4. Digman MA, et al. The phasor approach to fluorescence lifetime imaging analysis. Biophys J. 2008;94(2):L14–6.

    Article  CAS  Google Scholar 

  5. Klemm M, et al. FLIMX: a software package to determine and analyze the fluorescence lifetime in time-resolved fluorescence data from the human eye. PLoS One. 2015;10(7):e0131640.

    Article  Google Scholar 

  6. Sauer L, et al. Monitoring foveal sparing in geographic atrophy with fluorescence lifetime imaging ophthalmoscopy – a novel approach. Acta Ophthalmol. 2018;96(3):257–66.

    Article  Google Scholar 

  7. Peters S, Griebsch M, Klemm M, Haueisen J, Hammer M. Hydrogen peroxide modulates energy metabolism and oxidative stress in cultures of permanent human Muller cells MIO-M1. J Biophotonics. 2017;10(9):1180–8.

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Ophthalmology and Department of Clinical Research, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

    Chantal Dysli

  2. University of Utah, John A. Moran Eye Center, Salt Lake City, UT, USA

    Lydia Sauer

  3. Institute of Biomedical Engineering and Informatics, Technische Universität Ilmenau, Ilmenau, Germany

    Matthias Klemm

Authors
  1. Chantal Dysli
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  2. Lydia Sauer
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  3. Matthias Klemm
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Corresponding author

Correspondence to Chantal Dysli .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology and Department of Clinical Research, Inselspital Bern University Hospital, University of Bern, Bern, Switzerland

    Martin Zinkernagel

  2. Department of Ophthalmology and Department of Clinical Research, Inselspital Bern University Hospital, University of Bern, Bern, Switzerland

    Chantal Dysli

Supplementary Material

Supplementary Material

FLIO Image Acquisition Protocol

Subjects

  • Maximal dilated pupil (at least 6 mm diameter)

  • Measure both eyes (internal comparison)

Measurement

Preparation

  • Complete darkening of the room (switch off light, close doors, lower blinds, reduce light on computer screen)

  • Move computer screen away from subjects face

  • No angiography prior to FLIO (at least 2 days, better 1 week)

  • No use of topical fluorescein before FLIO measurement

Before Image Acquisition

  • Place subjects head correctly

  • Select Tau mode on touch panel under “more”

  • Use internal fixation light if possible, otherwise use external fixation light

  • Standard imaging field: centered on the fovea, 30° field of view

  • Optimize illumination of the fundus

  • Adjust focus on small/medium sized vessels on infrared (IR) image

  • After switching on the laser: acquire a single IR image, then start the measurement

During Measurement

  • Check head position (forehead and chin attached to the device)

  • Motivate subject to open the eyes widely, to look at the fixation target, and to keep the position of the head

  • Ensure maximal and uniform illumination of the fundus

  • Slight adjustment of position during measurement is possible by moving the camera head in the opposite direction of shadows in the live acquisition image: left/right, up/down; NO movement towards or away from subject

  • Leave filter as deep inside as possible (deep = lower filtering); on red warning (too many photons) move filter out stepwise

End of Acquisition

  • Both channels: minimum of 1000 photons in macula or at the darkest/slowest pixel (use “at cursor” function/select with mouse pointer) corresponds to approximately 1200–1500 photons in average

  • Measurement time: about 1.5 to 2 min/eye in dilated pupils

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Cite this chapter

Dysli, C., Sauer, L., Klemm, M. (2019). Image Acquisition and Analysis. In: Zinkernagel, M., Dysli, C. (eds) Fluorescence Lifetime Imaging Ophthalmoscopy. Springer, Cham. https://doi.org/10.1007/978-3-030-22878-1_6

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  • DOI: https://doi.org/10.1007/978-3-030-22878-1_6

  • Published:

  • Publisher Name: Springer, Cham

  • Print ISBN: 978-3-030-22877-4

  • Online ISBN: 978-3-030-22878-1

  • eBook Packages: MedicineMedicine (R0)

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