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本文报道利用异硫氰酸荧光素标记的花生凝集素(FITC-PNA)定量测定小鼠胸腺细胞表面的花生凝集素受体。并比较系统地研究了细胞数和 FITC-PNA浓度的关系、保温时间及比色的条件,从而确定荧光定量分析的最适条件。结果:胸腺细胞(8×10~6/ml)可结合16.6±3.7微微克分子数的PNA。本文亦对小鼠胸腺和脾脏细胞的荧光定量与荧光定性测定进行了比较。实验结果提示,荧光定性技术可在细胞水平反映形态学和数量的特征,而荧光定量分析法具有重复性好、灵敏度高、操作简便和更为客观、准确等优点。
This paper reports the quantitative determination of peanut lectin receptors on mouse thymocytes using fluorescein isothiocyanate labeled peanut agglutinin (FITC-PNA). The relationship between cell number and FITC-PNA concentration, incubation time and colorimetric conditions were systematically studied to determine the optimum conditions for quantitative fluorescence analysis. Results Thymocytes (8 × 10 ~ 6 / ml) could bind 16.6 ± 3.7 picograms of PNA. In this paper, fluorescence quantitative and fluorescent qualitative determinations of thymus and spleen cells in mice were also compared. The experimental results suggest that the fluorescence qualitative technique can reflect the morphological and quantitative characteristics at the cellular level, while the fluorescence quantitative analysis method has the advantages of good repeatability, high sensitivity, simple and convenient operation, more objective and accurate.