Liang J N
Howe Laboratory of Ophthalmology, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston 02114.
Curr Eye Res. 1990 May;9(5):399-405. doi: 10.3109/02713689008999604.
Tryptophan (trp) and non-tryptophan (non-trp) fluorescence measurements of the sectioned lenses were made using front surface technique. Measurements on concentrated protein solutions showed that inner filter effect (attenuation of both excited and emitted light) rendered the conventional right angle detection less accurate. For fluorescence measurement of the intact lens, because of the presence of a high protein concentration and abundant aggregated and insoluble proteins in the aged lens, a better result can be obtained by front surface detection. In this study, dissected human lenses were measured with front surface detection to observe age related changes. A decrease in trp fluorescence and an increase in non-trp fluorescence were observed with age. The emission maximum of trp fluorescence was shifted to a longer wavelength in old lenses. These results indicate that front surface fluorometry is more useful in studying proteins in concentrated solutions and intact tissues than the conventional right angle fluorescence detection.
采用前表面技术对切片晶状体进行色氨酸（trp）和非色氨酸（non-trp）荧光测量。对浓缩蛋白质溶液的测量表明，内滤光效应（激发光和发射光的衰减）使传统的直角检测不够准确。对于完整晶状体的荧光测量，由于老化晶状体中存在高蛋白浓度以及大量聚集和不溶性蛋白质，通过前表面检测可获得更好的结果。在本研究中，采用前表面检测对解剖后的人晶状体进行测量，以观察与年龄相关的变化。随着年龄增长，观察到色氨酸荧光降低，非色氨酸荧光增加。在老年晶状体中，色氨酸荧光的最大发射波长移向更长波长。这些结果表明，与传统的直角荧光检测相比，前表面荧光测定法在研究浓缩溶液和完整组织中的蛋白质时更有用。
