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目的测试A549细胞经过光镜及电镜制样处理后的收缩系数,为定量研究A549细胞及其超微结构奠定基础。方法对A549细胞分别进行涂片HE染色、涂片巴氏染色、石蜡包埋3μm切片HE染色及透射电镜制样半薄(3μm)切片HE染色处理。通过光学显微镜获取细胞图像,用Image-Pro Plus图像分析软件测试细胞的平均直径,然后以A549培养之等渗重悬细胞之平均直径为基准计算细胞的收缩系数。结果A549培养之等渗重悬细胞的平均直径为14.93μm。经涂片HE染色、涂片巴氏染色、石蜡切片HE染色和透射电镜制样半薄(3μm)切片HE染色处理后的A549细胞的平均直径分别9.88μm、9.98μm、10.29μm和10.59μm。以等渗重悬细胞之平均直径为基准,经涂片HE染色、涂片巴氏染色、石蜡切片HE染色和透射电镜半薄切片HE染色之细胞的收缩系数分别为0.66、0.67、0.69和0.71。结论经涂片HE染色、涂片巴氏染色、石蜡切片HE染色和透射电镜制样半薄切片HE染色处理后,A549细胞收缩明显,直径约收缩了1/3。因此,在实际测试中,欲知细胞收缩前之结构大小,须据细胞收缩程度进行必要的校正。
Objective To test the contraction coefficient of A549 cells treated by light and electron microscopy, and lay the foundation for the quantitative study of A549 cells and their ultrastructure. Methods A549 cells were stained with hematoxylin and eosin (HE) staining, smear Papanicolaou staining, paraffin embedded 3μm sections HE staining and transmission electron microscopy (HE staining) of semi-thin (3μm) sections. Cell images were taken by light microscopy and the cells were tested for average diameter using Image-Pro Plus image analysis software and cell contraction coefficients were calculated on the basis of the average diameter of isobaric suspended cells grown in A549. Results The average diameter of isobaric resuspended cells cultured in A549 was 14.93μm. The average diameter of A549 cells stained with Hematoxylin and Eosin (HE) staining, smear Papanicolaou, paraffin section HE staining and transmission electron microscopy showed that the average diameters of A549 cells after HE staining were 9.88μm, 9.98μm, 10.29μm and 10.59μm, respectively. The contractile coefficient of cells stained with hematoxylin and eosin (HE) staining, smear Papanicolaou, paraffin section HE and semi-thin section HE staining were 0.66, 0.67, 0.69 and 0.71 . Conclusion HE staining of the smear, smear Papanicolaou, paraffin sections HE staining and transmission electron microscopy semi-thin section HE staining, A549 cells shrink significantly, the diameter of about 1/3 contraction. Therefore, in actual tests, to know the size of the structure before the cells shrink, the necessary corrections must be made according to the degree of contraction of the cells.