Antibiotics Experiment - Cell sorting

Flow cytometry data on mouse fecal bacteria incubated for three hours with (or without) 20 uM 5-ethnyl-2'-deoxyuridine (EdU), a thymidine analogue, in a 1:1 ratio of reduced Brain Heart Infusion (rBHI) broth and fecal slurry (FS), to capture the actively replicating bacteria. <br> Bacteria were exposed to Polymyxin B (all samples with "P"), Vancomycin ("V"), or the vehicle (DMSO; "C") for one hour before incubation with EdU. The antibiotics had a final concentration of 0.1 mg.mL-1. <br> The bacterial cells underwent a click reaction with Alexa Fluor 647 azide (APC channel) to make newly replicated bacteria fluoresece. They were also stained with Sybr Green I nucleic acid stain (FITC channel) to capture all bacteria.

本数据集为小鼠粪便细菌的流式细胞术检测数据：将小鼠粪便细菌置于还原脑心浸液肉汤（reduced Brain Heart Infusion, rBHI）与粪便浆液（fecal slurry, FS）按1:1体积比混合的体系中，以添加（或不添加）20 μM 5-乙炔基-2'-脱氧尿苷（5-ethnyl-2'-deoxyuridine, EdU，一种胸苷类似物）的体系孵育3小时，以此捕获活跃增殖的细菌。
在进行EdU孵育前，细菌先经多粘菌素B（所有标注“P”的样本）、万古霉素（标注“V”的样本）或溶媒二甲基亚砜（DMSO；标注“C”）处理1小时，上述抗生素的终浓度均为0.1 mg·mL⁻¹。
细菌细胞与Alexa Fluor 647叠氮化物（APC通道）进行点击化学反应，使新增殖的细菌发出荧光；同时使用Sybr Green I核酸染料（FITC通道）对所有细菌进行染色，以标记全部待测细菌。