血球计数板检测破壁灵芝孢子粉破壁率方法的优化研究

Optimizing the hemocytometer measurement method for determining the sporoderm-broken rate of Ganoderma lucidum spore powder

中文关键词:  血球计数板  破壁灵芝孢子粉  破壁率  细胞染色技术

英文关键词:hemocytometer measurement  sporoderm-broken Ganoderma lucidum spore powder  sporoderm-broken rate  cell staining technology

中文摘要:

      目的 对现行常用的血球计数板检测破壁灵芝孢子粉破壁率的方法进行优化研究, 以降低检测误差, 避免人为主观判断, 并提高检测效率。方法 通过考察不同的悬浮液体系, 筛选出能使破壁灵芝孢子粉分散性最佳的悬浮液体系; 通过考察常用的细胞染色剂, 筛选出能是破壁灵芝孢子粉染色, 而灵芝孢子粉不被染色的最适染色剂。结果 优化后检测方法重复性与再现性良好, 重复性与再现性相对标准偏差(relative standard deviationg, RSD)均为0.3%; 回收率良好, 不同破壁率水平的样品回收率平均值在98.9%~100.9%范围内; 检测效率明显提高, 单个样品的检测时间从18.2 min降低至8.1 min。结论 本研究通过优化悬浮液体系及引用细胞染色技术与血球计数法结合, 降低了检测方法误差, 避免了人为主观判断, 建立了一种更为准确、客观、高效的破壁率检测方法, 为破壁灵芝孢子粉破壁率检测提供了可靠手段。

英文摘要:

      Objective To optimize the commonly used hemocytometer measurement method for deteimining the sporoderm-broken rate of Ganoderma lucidum spore powder, reduce experimental error, avoid subjective judgment by technician and improve the detection efficiency. Methods By examining different suspension systems, the best one was selected that could fully disperse the sporoderm-broken Ganoderma lucidum spore powder. By examining commonly used cell staining agents, the most suitable staining agent was selected that could stain broken Ganoderma lucidum spore powder while not staining Ganoderma lucidum spore powder. Results The optimized detection method had good repeatability and reproducibility, with a relative standard deviation (RSD) of both repeatability and reproducibility were 0.3%. The recovery rate was good, and the average recovery rate of samples with different levels of sporoderm-broken rate were within the range of 98.9% to 100.9%. The detection efficiency had been significantly improved, and the detection time for a single sample had been reduced from 18.2 min to 8.1 min. Conclusion The article optimizes the suspension system and combines cell staining technology with hemocytometer measurement to reduce detection method errors, avoid subjective judgments and establish a more accurate, objective and efficient method for detecting sporoderm-broken rate of Ganoderma lucidum spore powder, providing a reliable means for sporoderm-broken rate detection of Ganoderma lucidum spore powder.