Diff-Quik染色法在中华鳖血细胞染色中的应用

Application of Diff-Quik stain for the blood cells of Chinese soft-shelled turtles,Pelodiscus sinensis

采用Diff-Quik染色法对中华鳖的血涂片进行染色,用DP-71显微成像系统采集血片细胞显微图像,经Image-proExpress5.1测量和分析中华鳖各类血细胞大小、比例。结果表明:①Diff-Quik染色后中华鳖各类血细胞核质染色差别明显,核质界面明显,各类白细胞的颗粒明显、易分辨。②中华鳖红细胞、嗜中性粒细胞、嗜酸性粒细胞、嗜碱性粒细胞大小分别为17.86μm×12.43μm、16.26μm×13.67μm、12.28μm×10.98μm、14.63μm×12.83μm。嗜中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞占白细胞百分比分别为54.7、13.72、6.22和24.72,嗜碱性粒细胞仅为0.65。③比较Diff-Quik法、Giemsa法、Wright-Giemsa法对染色效果的影响,3种染色方法在细胞大小及白细胞分类比例上无明显差异。从核质界面、细胞颗粒特征性染色等来看,Diff-Quik法和Giemsa法优于Wright-Giemsa法。Diff-Quik法染色快捷、核质和颗粒染色清晰度更高,是一种适合大量标本快速染色的染色法,可适用于中华鳖的血细胞染色。

The blood smears of Chinese soft-shelled turtle,Pelodiscus sinensis,were used for hematologic observation after stained with Diff-Quik stain kit.The blood cell images were captured and analyzed with DP-71 digital imaging system and Image-pro Express 5.1 for the size measuring and blood cells sorting.The results showed：（1） The blood cells of Chinese soft-shelled turtle could be clearly distinguished by Diff-Quik stain both for nuclear and cytoplasm and for unique cytoplasmic particles of white blood cells.（2） The size of erythrocyte,neutrophils,eosinophils,basophils were 17.86 μm×12.43 μm,16.26μm×13.67 μm,12.28 μm× 10.98 μm,14.63 μm×12.83 μm,respectively.The percentage of neutrophils,eosinophils,monocytes,lymphocytes is 54.7%,13.72%,6.22%,24.72%,respectively.（3）No significant differences were observed among Diff-Quik stain、Giemsa stain and Wright-Giemsa stain in blood cell characterization and leukocyte counting.Diff-Quik stain and Giemsa stain showed better results than Wright-Giemsa stain both in nuclear and cytoplasm staining and in easier distinguished particles for different white blood cells.The Diff-Quik stain,giving better definition for nucleic and particles with more convenient and faster processing,could be considered a better method for blood cell stain for Chinese soft-shelled turtle,specially for large specimens staining.