分光光度法与显微计数法测定微小绿藻生物量的比较研究  

Methodological Comparison Between Spectrophotometry and Microscopic Counting in Measuring Biomass of Micro-Green Algae

作　　者：涂波 曹敏[1] 黄茜[1] 杨晓红[1] 

机构地区：[1]南方山地园艺学教育部重点实验室/西南大学园艺园林学院,重庆400716

出　　处：《西南大学学报（自然科学版）》2014年第8期44-50,共7页Journal of Southwest University(Natural Science Edition)

基　　金：重庆市科委项目(CSTC;2011AB1028)基金资助

摘　　要：使用紫外-可见分光光度计对6株绿藻的培养液进行600-800nm的波长扫描,在685nm处获得一个单纯显著的特征吸收峰.将6株微小绿藻藻液分别稀释成5个梯度,同步进行血球计数板显微计数和紫外-可见分光光度计OD685值测定.稀释藻液的分光光度法拟合曲线R2≥0.990,显微计数法拟合曲线R2≥0.940,OD685计数R2在0.914与0.989之间;同步培养6株绿藻25d,从第6d开始,每天650-700nm扫描藻液测量OD,确认吸收峰是否会发生偏移,并同步测定藻液细胞浓度和藻液OD685值,比较拟合结果.纯种验证试验中,单纯特征吸收峰均在685nm处再现.OD685显微计数散点拟合R2在0.780与0.970之间,误差较大的2个藻株是蹄形藻和针形纤维藻;将6株藻培养20d的藻液混合,并稀释成浓度梯度,测量OD685与显微计数,混合藻液OD685计数R2=0.972.混合藻种验证试验显示出OD685计数对纯种和混合藻种都有着良好的精确度.试验证明,微小绿藻在600-800nm波段的最大特征吸收峰稳定地发生在685nm处.OD685值能够准确反映藻液中纯种或混合微小绿藻的生物量或生物量的变化,精确度和稳定性高于血球计数板显微计数.UV-Vis spectrophotometer was used to scan the culture solution of 6 strains of green algae be- tween 600 nm and 800 nm of wavelength. The micro-green algae were diluted into 5 concentration gradients. A special single characteristic peak appeared at 685 nm of wave length in all the 6 strains. Concentration and OD value of the algae liquors were measured separately with a hemacytometer under microscope and with a UV-Vis spectrophotometer scanned at 685 nm at the same time. The linear fit degree for the two methods was very high. To further verify the above results, the 6 strains of algae were synchronously cultivated for 25 days. From the 6th day, the algae solutions were scanned every day between wavelength 650-700 nm in order to further confirm whether the characteristic absorption peaks would offset or not. The concentration and OD68s value of the algae solutions were synchronously measured every day. The linear fit results of the verification test and the compound algae solutions linear fit results were compared. The 6 strains of algae were also cultivated 20 days in liquid medium. Then they were mixed all in the same volume and diluted into 5 concentration gradients. The concentration and OD685 value of the mixed algae solutions were also measured. Again, the special single characteristic absorption peak appeared at 685 nm. R2 of the algae solutions linear fit were 90. 990. R2 of microscopic counting scatter linear fit were 0. 940. OD685 counting R2 was between 0. 914 and 0. 989. After cultivating 25 days, the absorption peak of microgreen algae also appeared at 685 nm and R2 was from 0. 780 to 0. 970. The mixed algae solution linear fit R2 was 0. 972. All the data above clarified that OD685 value could accurately present the biomass or the biomass change of micro-green algae with R^2 〉0. 990, and that spectrophotometry is more accurate and stable than microscopic counting.

关 键 词：微小绿藻 生物量 显微计数 光谱计数 特征吸收峰 

分 类 号：Q939.9[生物学—微生物学]