Rose Bengal sodium is a potent inhibitor of VGlut and vesicular monoamine transporter (VMAT) (Ki of 19 and 64 nM for VGlut andVMAT, respectively）

使用方法：
a.溶液配制：
1.将Rose Bengal sodium溶于DMSO中配置成10mM的储存液，注意避光，请将储存液分装并放在-20℃或者-80℃，避免反复冻融。
2.使用时母液稀释成一定梯度的工作液备用。
b.操作步骤：
1.将细菌培养至 10^8 CFU/mL 浓度，然后用无菌生理盐水连续十倍稀释至 10^ 3-10 ^7 CFU/mL 浓度。
2.将 3 mL细菌悬浮液转移到直径为 2.5 cm的平底 20 mL小瓶中。在实验之前，从每个小瓶中抽取样本以确定初始细菌浓度。
3.除对照组外，在所有小瓶中加入不同浓度的 Rose Bengal sodium。加入 Rose Bengal sodium 后，所有后续操作均在黑暗条件下进行。
4.将含有或不含 Rose Bengal sodium 的细菌悬浮液的小瓶放在 VU03H 塑料支架的超声槽底部，用频率为 38 kHz、电场强度为 4.1 W/cm^3 的超声处理 1 至 10 分钟。
5.处理完成后，将 100 μL 的样品用十倍稀释稀释，铺在 BHA 板上。平板在 37°C 下孵育过夜，使用活菌计数法确定细菌细胞浓度。

Bacteria were grown to a concentration of 10^8 CFU/mL and then diluted with sterile saline by serial decimal dilutions to concentrations of 10^3–10^7 CFU/mL.?Three milliliters of bacterial suspensions was transferred into flat-bottom 2.5 cm diameter 20 mL vials.?Before the experiment, samples were taken from each vial to determine the initial bacterial concentration.?Free Rose Bengal at various concentrations or Rose Bengal immobilized onto silicone was added to all vials, except for the controls.?After adding Rose Bengal, all subsequent procedures were performed under dark conditions.?Vials with bacterial suspensions with or without Rose Bengal were held tight to the bottom of an ultrasonic bath in a plastic holder VU03H ?and treated by ultrasound at a frequency of 38 kHz and a field strength of 4.1 W/cm^3 for 1 to 10 min. After the treatment, 100 μL samples were diluted by several decimal dilutions and were spread over BHA plates with a Drigalsky spreader.?The plates were incubated at 37 °C overnight and the bacterial cell concentration was determined taking dilutions into account using the viable count method, where CFU were counted by means of a colony counter Scan 500.?In control experiments, bacterial cultures were tested in the absence of Rose Bengal without sonication, in the absence of Rose Bengal under sonication and in the presence of Rose Bengal without sonication[1].