应用流式细胞术测定淡水、海洋沉积物中异养细菌前处理条件的选择

目前流式细胞术(FCM)已被广泛应用于水体异养细菌的检测,采用适当的前处理方法可将黏附在沉积物颗粒上的异养细菌提取到水相中进而利用FCM进行检测.选择合适的前处理方法是将FCM应用于沉积物异养细菌检测的关键.论文对FCM测定沉积物中异养细菌的前处理方法进行了探讨,实验同步考虑了淡水及海洋沉积物.结果表明,对于实验所用淡水、海洋沉积物,较优的前处理条件为:1mmol·L-1焦磷酸钠作为分散剂,暗处孵育10min,20w、40KHz水浴超声1min,并每30s人工振荡1次,2800r·min-1常温离心萃取3次.

Optimization in Pretreatment Conditions for Determination of Heterotrophic Bacteria from Freshwater and Marine Sediments Using Flow Cytometry

The Flow Cytometry (FCM) has been widely used to determine the heterotrophic bacteria in water environment presently. Also, it can be used to determine the heterotrophic bacteria in benthos environment after the appropriate pretreatment. The treatment of extracting sediment-attached bacteria to water environment is the key point of applying the FCM to the study of benthos. This paper discussed the methods of pretreatment for the FCM to detect the heterotrophic bacteria both in freshwater and marine sediments. Results showed that the relatively preferable pretreatment conditions for sediments were: sodium pyrophosphate as the dispersant with the final concentration of 1mmol·L-1 and incubated 10min in dark; sonicate 1min in water bath at 20w, 40KHz power with a manual shake every 30s; and centrifugal extraction 3 times with 2800 r·min-1 at room temperature.