fsc_files_Fc7_Salmon

fsc files and metadata for analysis This repository contains the raw fsc files for the experiment investigating the effect of phage therapy and antibiotics on treating Flavobacterium columnare Fc7 infections in Atlantic salmon fry. The bacterial communities in the water were sampled (1 mL) right before and immediately after adding Fc7 and treatment. Throughout the experiment, the water was sampled after treatment and water exchanges were performed at 0.5, 1, 2, 4, 6, 8 and 10 DPI. The water samples were fixated through a 15-minute incubation in 0.1% Glutaraldehyde before being snap-frozen using liquid nitrogen. The bacterial density was quantified using flow cytometry (Attune NxT, ThermoFisher). Data were collected using the blue laser (488 nm) with detection in BL1 and BL3 using a BL1 threshold of 1500-3000 (depending on the sample). Instrument voltages were FSC 320V, SSC 260V, BL1 320 V and BL3 350V. Samples were acquired by running 160 uL sample at a 100 uL/min flowrate. Samples were vortexed before being sampled and the zip was cleaned between approximately every 6th sample. The water samples were stained with SYBR green I (2x) which emits a green fluorescent signal when it binds to DNA. Samples were incubated for 15 minutes in dark at 37 degrees with the stain before analysis. Samples were diluted in 0.2 um filtered PBS (1x) to obtain stable sample aquation and dilute background noise present in the sample. 0.2 um filtered PBS and 0.2 um filtered fish water were used as negative controls and the pure strains of Fc7 as positive controls.

用于分析的FSC文件及元数据。
本仓库收录了探究噬菌体疗法与抗生素联合治疗大西洋鲑鱼苗柱状黄杆菌Fc7感染效果的实验原始FSC文件。
实验过程中，在添加Fc7菌株及施加治疗前后即刻采集水体细菌群落样本（体积1 mL）；实验全程在施加处理后及各换水节点采集水样，换水节点设置为感染后第0.5、1、2、4、6、8及10天（Days Post Infection，简称DPI）。
水样采集后，先在0.1%戊二醛溶液中孵育15分钟进行固定，随后采用液氮快速冷冻。
细菌密度通过流式细胞术（Attune NxT型，赛默飞世尔科技）进行定量：使用488 nm蓝色激光激发，于BL1和BL3通道检测信号，BL1通道阈值设为1500~3000（依样本具体情况调整）；仪器电压参数为：前向散射光（Forward Scatter，简称FSC）320 V、侧向散射光（Side Scatter，简称SSC）260 V、BL1通道320 V、BL3通道350 V。
样本上机检测流程为：以100 μL/min的流速进样，进样体积为160 μL；采样前对样本进行涡旋混匀，每采集约6个样本后清洁取样装置。
水样采用2×浓度的SYBR Green I染料染色，该染料与DNA结合后可发出绿色荧光信号。染色完成后，将样本置于37℃、避光条件下孵育15分钟，随后进行上机分析。
为保证样本检测稳定性并降低背景噪音，样本采用经0.2 μm滤膜过滤的1×磷酸盐缓冲液（PBS）进行稀释。实验设置两类阴性对照：经0.2 μm滤膜过滤的PBS，以及经0.2 μm滤膜过滤的养殖水体；以Fc7菌株纯培养液作为阳性对照。