[Limits of the confocal laser-scanning technique in measurements of time-resolved autofluorescence of the ocular fundus].
OBJECTIVE
The 2-dimensional measurement of the time-dependent autofluorescence has in combination of confocal laser scanner technique and time correlated single photon counting the potential for the investigation of the metabolism state at the eye ground. It was to be examined, to what extent these measurements are influenced through the auto-fluorescence of the crystalline lens.


METHODS
The time-dependent auto-fluorescence was measured on eyes of 21 patients with age-related macular degeneration (AMD) and of 26 healthy subjects in 40 degrees fundus images. The experimental set-up used, was developed at the eye clinic of the University of Jena. In this scanning laser ophthalmoscope, the fundus was excited at 446 nm by pulses of 100 ps full width at half maximum and 40 MHz repetition rate. The auto-fluorescence was detected with a time resolution of 25 ps for wavelengths > 500 nm. The dynamic auto-fluorescence of each pixel was approximated by a tri-exponential model function. In order to determine the influence of the fluorescence of the crystalline lens, healthy eyes and eyes were compared with implanted artificial intra-ocular lenses. For comparison the frequency distributions were used, which the decay times t1, t2 and t3 in the fundus images had been calculated with.


RESULTS
Clear differences were shown in the frequency distributions of the fluorescence lifetimes between AMD-patients and healthy subjects, It resulted, however, from a crucial analysis that the fluorescence of the crystalline lens is not suppressed by the confocal laser scanning principle sufficiently. In particular the long lifetime t3 is overlapped by the fluorescence of the crystalline lens of about 4.6 ns. To the complete suppression of the influence of the lens fluorescence in measurements of the time-resolved fundus auto-fluorescence it is suggested to combine the confocal laser scanning technique with the principle of the aperture diaphragm division.


CONCLUSIONS
In order to be able to determine the capability of measurements of the time-dependent fundus auto-fluorescence as diagnostic tool exactly, the influence of the crystalline lens is to reduce through the combination of confocal laser scanning ophthalmoscopy with the principle of the aperture diaphragm division diversely.
Schweitzer
D
D
Experimentelle Ophthalmologie, Augenklinik der Friedrich Schiller Universität Jena. Dietrich.schweitzer@med.uni-jena.de
Hammer
M
M
Schweitzer
F
F
D005451:Fluorescein Angiography / Q000379:methods*; D006801:Humans; D008268:Macular Degeneration / Q000473:pathology*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009887:Ophthalmoscopy / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity