Doughty M J, Blades K, Button N F, Wilson G
Department of Vision Sciences, Glasgow-Caledonian University, UK.
Ophthalmic Physiol Opt. 2000 Sep;20(5):391-400.
To assess whether the size of the cells obtained by conjunctival impression cytology can be quantitatively assessed by measurement of the longest dimension of the cells.
Under topical benoxinate anaesthesia, cells were removed from the normally exposed nasal bulbar conjunctival surface using a 0.4 micron pore diameter filter (Biopore filter; type Millcell-CM). The filters were stained with haematoxylin after ethanol denaturation, photographed at 40 x magnification, and 35 mm slides prepared. An optical overlay method was used to outline the borders of sets of 30-35 contiguous cells on each image. The cell area, longest and shortest dimensions were measured by planimetry to an accuracy of +/- 3%.
Analyses of 20 sets of samples, from individuals aged 21 to 48 years and without clinically significant ocular surface disease, revealed a median cell area of 133 micron 2 (n = 621, range 46-1602 micron 2; average 212 micron 2), a median longest dimension of 13.9 microns (range 6.6-68.8 microns; average 16.9 microns) and a shorter dimension of 10.0 microns (range 4.7-43.0 microns; average 11.9 microns); the distributions of values indicated bimodality. Most cells had a long:short ratio (L:S ratio) value between 1.00 and 1.80, but 11.9 +/- 6.1% of the cells had L:S ratios between 1.80 and 4.60. The overall relationship between the longest dimension and the area of the cells was nonlinear, with cells having larger L:S ratios having disproportionately smaller areas.
The superficial conjunctival cells are small, and their longest dimensions are systematically related to area. Analyses of cell shape indicate further possible ways of identifying different cells on the bulbar conjunctiva. Compared to literature values, there is a substantial overlap in longest dimensions of the conjunctival cells with those of cells that can be collected off the corneal surface. This means that superficial conjunctival and corneal cells cannot be distinguished simply on the basis of measurements of the long dimension.
评估通过测量结膜印迹细胞学所获细胞的最长直径,能否对细胞大小进行定量评估。
在丁氧卡因表面麻醉下,使用孔径为0.4微米的滤器(Biopore滤器;Millcell-CM型)从正常暴露的鼻侧球结膜表面获取细胞。滤器经乙醇变性后用苏木精染色,在40倍放大倍数下拍照,并制备35毫米幻灯片。采用光学叠加法在每张图像上勾勒出30 - 35个连续细胞组的边界。通过平面测量法测量细胞面积、最长和最短直径,测量精度为±3%。
对20组样本进行分析,样本来自年龄在21至48岁且无临床显著眼表疾病的个体,结果显示细胞面积中位数为133平方微米(n = 621,范围46 - 1602平方微米;平均212平方微米),最长直径中位数为13.9微米(范围6.6 - 68.8微米;平均16.9微米),较短直径为10.0微米(范围4.7 - 43.0微米;平均11.9微米);数值分布呈双峰性。大多数细胞的长宽比(L:S比)值在1.00至1.80之间,但11.9 ± 6.1%的细胞L:S比在1.80至4.60之间。细胞最长直径与面积之间的总体关系是非线性的,L:S比越大的细胞面积相对越小。
表层结膜细胞较小,其最长直径与面积存在系统性关联。细胞形状分析表明,在球结膜上识别不同细胞可能还有其他方法。与文献值相比,结膜细胞的最长直径与可从角膜表面采集的细胞的最长直径有很大重叠。这意味着不能仅根据长径测量来区分表层结膜细胞和角膜细胞。