图像分析与人工计数视网膜神经节细胞方法的比较

Comparison of image analyzer-aided and conventional retinal ganglion cell counting method

目的 探讨图像分析仪计数视网膜神经节细胞 (retinal ganglion cell,RGC)的特点和实际应用的可能性。 方法 自眶内切断 18只成年大鼠左眼视神经 ,近侧断端留置浸有 5 %荧光金的明胶海绵。分别于手术后 2、7、14 d处死动物 ,每一时间点各 6只。取左眼视网膜经后固定平铺于载玻片上 ,荧光显微镜下分别以人工视网膜分区取样估测法和 Quantimet5 70图像分析仪计数荧光金逆行标记的 RGC。 结果 RGC数目随手术后存活时间延长而下降 ,手术后 2、7、14 d图像分析仪和人工计数法计数的 RGC总数分别为 (12 0 6 6 3± 90 89)、(5 92 85± 170 71)、(1780 2± 1984 )个 / mm2 和 (1182 37± 7898)、(5 76 4 8±14 5 33)、(180 70± 14 6 1)个 / mm2 ,同一时间点两组间计数结果差异无显著性的意义 (P>0 .0 5 )。 结论 图像分析仪计数 RGC具有较为简便、客观和可重复计数的特点 ,可用于计数 RGC的实验研究。

Objective To investigate the characteristics and possibility of using an image analyzer-aided method to count axotomized retinal ganglion cells (RGCs). Methods The left optic nerves of 18 rats were transected intraorbitally and a piece of gelform soaked in 5% fluorogold was applied to the ocular stump to retrogradely label the surviving RGCs. All animals were executed 2, 7 or 14 days after the operation (n=6 for each time point), respectively. The left retinae were removed, post-fixed and whole-mounted on the slides. The numbers of labeled RGCs were counted using both the conventional sampling method and image analysis, and compared statistically between the two methods. Results The number of surviving RGCs decreased sharply [(12 0663±9 089), (59 285±17 071) and (17 802±19 84) cells/mm 2 for image analyzer-aided method, and (118 237±7 898), (57 648±14 533) and (18 070±1 461) cells/mm 2 for conventional sampling method] when the survival time increased from 2 to 7 and 14 days. No significant difference was detected between the two groups at any corresponding time points. Conclusion The image analyzer-aided method is convenient, objective and reproducible, which can be used in the studies where counting RGCs is needed.