In vivo measurement of time-resolved autofluorescence at the human fundus.
An experimental setup for measurement of time-resolved autofluorescence of the human eye fundus is demonstrated. The method combines laser scanning technique and time-correlated single photon counting. The light source is a laser diode, delivering pulses of about 100 ps duration at a repetition rate of 40 MHz. The excitation wavelength is 446 nm and the cutoff wavelength of fluorescence detection is at 475 nm. The autofluorescence can be determined with a spatial resolution of 80 x 80 microm2 and 25 ps time resolution. The fluorescence decay is optimally approximated by a biexponential model. The dominating lifetime tau1 is shortest in the macula (320 to 380 ps) and reaches 1500 ps in the optic disk. The lifetime tau2 varies between 2 ns and 5 ns, but the spatial distribution is more homogeneous. Respiration of 100% oxygen for 6 min leads to changes in the fluorescence lifetime pointing to detection of coenzymes. Diagrams of lifetime tau2 versus tau1 are well suited for comparison of substances. Such lifetime clusters of a 20 deg macular field of a young healthy subject and of a patient suffering from dry age-related macular degeneration overlap only partially with tau2-tau1 clusters of lipofuscin.
Schweitzer
Dietrich
D
University of Jena, Department of Experimental Ophthalmology, Bachstr. 18, 07740 Jena, Germany. Dietrich.Schweitzer@med.uni-jena.de
Hammer
Martin
M
Schweitzer
Frank
F
Anders
Roswitha
R
Doebbecke
Torsten
T
Schenke
Stefan
S
Gaillard
E R
ER
Gaillard
E R
ER
D000465:Algorithms; D005451:Fluorescein Angiography / Q000379:methods*; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008268:Macular Degeneration / Q000175:diagnosis*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009887:Ophthalmoscopy / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity