Q：做肝脏巨噬细胞清除需要灌流，还有流式有没有什么建议？

A：没有灌注，流式毫无意义，因为巨噬细胞没有分到，怎么看差异。流式建议检测F4/80和CD11b。推荐尾静脉注射，24h后取肝脏，做免疫组化。免疫组化不用灌流。

Q：体内巨噬细胞清除剂空白脂质体对照这个是单室脂质体还是多室脂质体？

A: 多室的。

Q：巨噬细胞清除剂的浓度是多少？

A: 5mg/mL。

Q: Clodronate Liposomes(From Vrije Universiteit Amsterdam) 体内巨噬细胞清除剂这个产品的脂质体粒径是多大?

A: 150nm~3um.

Q：收到 Clodronate liposomes 后该如何保存 操作？

A：收到氯磷酸脂质体后，如果不能马上使用，需放置在 4℃冰箱。禁止冻存！使用时作为原液使用， 不能进行稀释。脂质体很容易发生沉淀，建议使用之前轻轻混匀，另外 4℃的氯磷酸脂质体不能马上使用，需要恢复到室温才能进行注射。

Q：Clodronate liposomes 能否用于体外巨噬细胞清除？

A：产品可以用于体外巨噬细胞的清除，但是这种方法比较适合体内实验。主要是由于在体外培养过程中，从死亡细胞释放的氯磷酸或者是从脂质体中“渗漏”出来氯磷酸，会一直存在于培养基中，但是在体内，氯磷酸的半衰期很短，很快就会被肾脏清除。虽然游离的氯磷酸不会进入细胞也不会进入脂质体，但是一旦他们在培养基中积累就会慢慢地进入细胞中。

Q：在静脉注射 Clodronate liposomes 后动物很快就死亡了，是什么原因？

A：一方面有可能是动物注射了不均一的脂质体悬液。建议在使用之前轻轻摇晃混匀。脂质体在体外   一段时间后会发生沉降。如果在很长一段时间内需要一次注射多只动物，那么脂质体便会在注射器中   发生沉淀，形成不均一悬液，这样一来，第一只注射的计量与最后一只注射的计量就会发生明显的变   化。另外一方面有可能是脂质体刚从冰箱取出后，没有恢复到室温。

Q：对于清除不同器官或是组织内的巨噬细胞应该注射什么位置，注射剂量多少，注射时间和周期如何确定？

A：这个要涉及到具体的实验方案设计，建议客户自行设计实验方案，寻找符合自己实验目的的文献进行参考。

Q：动物在注射 Clodronate liposomes 后几天就死亡了？

A：这个可能是操作过程中受到细菌的污染，体内清除巨噬细胞后会增加例如病毒粒子 细菌或者酵母感染的几率。

Q：静脉注射 Clodronate liposomes 后大鼠脾脏/肝脏中的 ED1+细胞并没有完全消失？

A：大鼠中成熟的巨噬细胞呈现 ED1 和 ED2 双阳性，但是有一些没有吞噬功能的或者吞噬作用小的前体细胞也是 ED1+，但是呈现 ED2-。因此，所有 ED2+的细胞都可以被完全清除，但是 ED1+的细胞却只能被部分清除，清除的比率取决于前体/成熟巨噬细胞的比例。

Q：Clodronate liposomes 没有达到期望的效果？

A：氯磷酸脂质体都是大批量的生产，每批次都会对氯磷酸的浓度以及一些可能的污染物进行检测，确保无误后才会销售给客户。另外，脂质体容易收到极端温度的影响。正确的运输和储存方式：4~8℃，悬液既不能被冻存，也不能被加热超过 30℃。自收到货物起，需要在 3 个月内使用完，不然有可能会影响使用的效果。

Q：在静脉注射时，能不能增加注射的体积？

A：静脉注射时需要依据动物的体重，不能超过 0.1ml/10mg，但是进行腹腔注射可以适当的增加注射的体积。皮下注射需要根据注射位点的容量进行决定。

Q: 巨噬细胞清除剂，运输的过程中产品冻上了,是不是不能用了？

A：是的，无法使用。

Q: 巨噬细胞清除剂是阴离子么，我看是脂质体包裹的，应该是负电吧？

A: 是的，阴离子的。

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