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[共焦激光扫描技术在测量眼底时间分辨自体荧光方面的局限性]

[Limits of the confocal laser-scanning technique in measurements of time-resolved autofluorescence of the ocular fundus].

作者信息

Schweitzer D, Hammer M, Schweitzer F

机构信息

Experimentelle Ophthalmologie, Augenklinik der Friedrich Schiller Universität Jena.

出版信息

Biomed Tech (Berl). 2005 Sep;50(9):263-7. doi: 10.1515/BMT.2005.038.

Abstract

GOAL

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.

MATERIAL AND METHOD

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.

CONCLUSION

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.

摘要

目标

结合共焦激光扫描技术和时间相关单光子计数,对随时间变化的自发荧光进行二维测量,具有研究眼底代谢状态的潜力。本研究旨在考察这些测量在多大程度上受到晶状体自发荧光的影响。

材料与方法

在40度眼底图像中,对21例年龄相关性黄斑变性(AMD)患者和26例健康受试者的眼睛进行了随时间变化的自发荧光测量。所使用的实验装置是由耶拿大学眼科诊所开发的。在这种扫描激光检眼镜中,眼底由半高全宽为100 ps、重复频率为40 MHz的脉冲在446 nm处激发。对于波长>500 nm的光,以25 ps的时间分辨率检测自发荧光。每个像素的动态自发荧光由三指数模型函数近似。为了确定晶状体荧光的影响,将健康眼睛和植入人工晶状体的眼睛进行了比较。为了进行比较,使用了频率分布,其中眼底图像中的衰减时间t1、t2和t3已经计算出来。

结果

AMD患者和健康受试者之间荧光寿命的频率分布显示出明显差异。然而,关键分析表明,共焦激光扫描原理对晶状体荧光的抑制作用不够充分。特别是长寿命t3与约4.6 ns的晶状体荧光重叠。为了在时间分辨眼底自发荧光测量中完全抑制晶状体荧光的影响,建议将共焦激光扫描技术与孔径光阑分割原理相结合。

结论

为了能够准确地确定随时间变化的眼底自发荧光测量作为诊断工具的能力,应通过将共焦激光扫描检眼镜与孔径光阑分割原理相结合,以不同方式减少晶状体的影响。

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