Department of Vision Sciences, Glasgow-Caledonian University, Glasgow, Scotland.
Eye Contact Lens. 2011 Mar;37(2):50-6. doi: 10.1097/ICL.0b013e31820c6df0.
To assess the use of a public domain software Image J (NIH Image, Bethesda, MD) to make dimensional measures of human bulbar conjunctival cells.
Impression cytology samples were obtained from the nasal bulbar conjunctiva, and color images were taken at 200× magnification and projected and an overlay prepared or the image uploaded into Image J. The final image magnification for the overlays was approximately 2× that on the computer screen. For either overlays or screen images, linear measures were made from 30 or 25 cells of the cell longest dimension (LONG) and of the longest dimension of the nucleus (NUCLONG). The predicted variability in measures, from the calculated average values for any particular image, was systematically assessed, and the overall average results were compared.
Image J measures were within ±2% agreement with overlays, with LONG and NUCLONG measures increasing with the grade of squamous metaplasia. The net difference in LONG measures was within -2.65 to + 2.93 μm, and NUCLONG measures were within -0.87 and +1.39 μm. There was a slight tendency for NUCLONG measures to be systematically overestimated on smaller nuclei when using Image J on the color images.
The manual use of Image J on on-screen images can provide reasonably accurate objective measures of bulbar conjunctival cells, as compared with a higher magnification manual overlay technique. This should be suitable for comparisons between samples and to assess the effects of any disease-related changes or any interventions.
评估使用公共领域软件 Image J（NIH Image，贝塞斯达，马里兰州）对人球结膜细胞进行二维测量。
从鼻球结膜获取印迹细胞学样本，在 200×放大倍数下拍摄彩色图像，并对图像进行投影和叠加，或者将图像上传到 Image J 中。叠加后的最终图像放大倍数约为计算机屏幕上的 2 倍。对于叠加图像或屏幕图像，从 30 或 25 个最长细胞维度（LONG）和最长核维度（NUCLONG）的细胞中进行线性测量。系统评估了从任何特定图像的计算平均值预测的测量值的可变性，并比较了总体平均值结果。
Image J 测量值与叠加值相差在±2%以内，LONG 和 NUCLONG 测量值随鳞状上皮化生程度的增加而增加。LONG 测量值的净差异在-2.65 至+2.93μm 之间，NUCLONG 测量值在-0.87 和+1.39μm 之间。当在彩色图像上使用 Image J 时，较小核的 NUCLONG 测量值存在系统高估的轻微趋势。
与更高放大倍数的手动叠加技术相比，在屏幕图像上手动使用 Image J 可以提供对球结膜细胞的合理准确的客观测量。这应该适合于样本之间的比较，并评估任何与疾病相关的变化或任何干预措施的影响。
