文献中的实验方法原文如下：
E. coli cultures were grown on LB agar (Difco, Sparks,
MD) for 2 days at 37 °C prior to experimentation. Cells were
subsequently transferred from the plate under sterile conditions
to 250 mL of phosphate buffer solution (PBS, pH 7.0).
Thesolutionwasserially diluted to the required concentration
range. Five milliliters of the solution was filtered in triplicate
for each dilution onto presterilized filter membranes (0.45-
ím mixed cellulose esters, 47-mm diameter (Millipore,
Pittsburgh, PA)). Following filtration, the membranes were
placed on sterile 10-mL Petri dishes containing 9 mL of LB
agar. The Petri dishes with the filter membranes in place on
the LB agar were exposed to the afterglow plume emitted
from the grounded hollow slot electrode for the required
time. After the plasma treatment, the Petri dishes were
incubated at 37 °Cfor 1 day prior to determining the resulting
number of colony forming units (CFU).
凭借我poor的英语知识搞成了汉语，大家凑合着看吧。
　　大肠杆菌在37摄氏度环境下，在实验开始前于LB琼脂培养基（Difco公司，Sparks，MD）中生长两天。随后将大肠杆菌细胞体从无菌板转移到250毫升的磷酸缓冲溶液中（PBS，PH 7.0）。将溶液持续稀释至要求的浓度范围内。再取三份五毫升上述溶液过滤，每份溶液都用使用直径47毫米混有纤维素酯的无菌过滤膜（0.45微米）（Millipore公司，Pittsburgh，PA）过滤。过滤后，将膜放在装有9毫升LB琼脂的容量为10毫升的无菌培养皿中。再将培养皿暴露在从接地中空槽状电极发出的等离子羽辉到规定时间，培养皿中过滤膜位于LB琼脂上。经过等离子体处理后，将这些培养皿在37摄氏度下培养一天，然后得出菌落形成单位（CFU）的结果。

那么我还有几个问题：
1. 0.45微米的膜能过滤细菌么。我听卖仪器的说要0.2微米才行。这里的细菌是纯种的大肠杆菌。
2. 滤膜处理后培养的问题，培养皿不是得倒过来培养么？ 而且就算不倒过来，滤膜的细菌如何接触到琼脂的？另外像这种情况下，过滤后细菌应该是很集中分布的，直接培养会不会很密集无法计数？
难道看英文文献也会被坑啊？各种看不懂。
坐等大神，如果有需要看原文的请留言，谢谢大家了！ 返回小木虫查看更多