流式细胞仪在活体微藻计数中的应用

Application of Flow Cytometry in Living Microalgae Count

通过对不同染色剂的不同浓度的不同微藻运用流式细胞仪计数方法,将其结果与用常规血球计数板计数结果比较,目的是探讨流式细胞仪计数的正确性与可靠性。结果:发现流式细胞仪的计数范围是1×10~2~1×10~6,较之公认的计数范围1×10~4~5×10~5要宽,且流式细胞仪计数在1×10~3~1×10~6的相对标准偏差(RSD)基本在2%左右,最大不超过5%。在1×10~2误差在15%左右。同时,实验还探讨了流式细胞仪对异弯藻和塔胞藻的混合悬液(n(A)∶n(B)=3∶1或1∶3)的计数。不管塔胞藻与异弯藻以何种比例混合,混合液的理论浓度与实际由流式细胞仪计数的浓度都很相近。而且两种不同混合藻液的相对标准偏差小于3%。结论是流式细胞仪对活体微藻计数结果是可靠的。

Methods of flow cytometry on different fluorescent dyes （ Viacount and PI ） are used to enumerate the different concentration of different viable microalgae. At the meantime, we also need compare the result of the rapid counting by cytometer to the result of conventional hemocytometer. The objective is to investigate the correctness and reliability of flow cytometry. As a result,It can be found that the counting range by cytometer is between 1 ×102 and 1 × 106. It~ counting range is wider than the current study has proved （from 1 ×104 to 5× 105） and the viable mieroalgae（ from 1 × 105 to 1 × 107）. The RSD of counting by cytometer is about 2% ,when enumerating rang from 1 × 103 to 1 ×106. And even the max RSD is less than 5%. The RSD is about 15% ,only when counting the concentration as low as 1 × 102. At the meantime,experiment also explored the en.umeration of mixed suspension by cytometry. The mixed suspension is constituted by Heterosignm akashiwo and Pyramimonas sp.. No matter what proportions it is,the enumeration and theoretical value is very close. And the RSD of the counting is less than 3%. There is a conclusion that the counting in living microalgae by cytometry is very reliable.