MitoTracker® Deep Red FM是一种细胞渗透型的carbocyanine-based的深红外红色荧光探针(Ex=644 nm,Em=665 nm),包含标记线粒体的弱巯基反应性的氯甲基官能团,只需简单孵育细胞,即可被动运输穿过细胞膜并直接聚集在活性线粒体上。一旦线粒体被染色后,还能根据后续实验的需求进行固定(醛类固定剂如甲醛)和透化(醛类去污剂如Triton X-100),探针依然维持在细胞内。本品适合双标实验,因其红色荧光与其他的绿色荧光探针具有良好的分辨率。
虽然传统的线粒体荧光探针如TMR和罗丹明123,也能很容易的聚集在功能线粒体上,但是一旦线粒体膜电位丧失即会被洗掉,从而在一些需要细胞进行醛类固定或者包含线粒体能量状态影响因子的实验中,使其应用大受限制。
CAS号(CAS NO.) | N/A |
分子式(Formula) | C34H36Cl2N2 |
分子量(Molecular weight) | 543.58 |
外观(Appearance) | Blue solid |
激发波长(Ex) | 644 nm |
发射波长(Em) | 665 nm |
结构式(Structure) | ![]() |
室温运输;-20℃避光干燥保存,有效期1年。
Q:染色只能看到在细胞核周围有红色的弥散状,看不到线粒体的结构?
A:可能是探针浓度太高或者探针孵育后没有清洗干净,造成有非特异性染色。
Q:活细胞内直接做线粒体染色的试剂有哪些?
A:40740、40741、40742、40743 都可以。
Q:没有 HBSS 用什么缓冲液,加多少体积上流式?
A:可以使用 PBS 或是无血清培养基替代,添加体积足够做流式即可 100-200ul。
Q:染色结果可以使用多功能酶标仪观察吗?
A:可以,最好是使用荧光显微镜或是共聚焦显微镜、流式仪器。
Q:线粒体探针的保质期是多久?
A:按规定标准储存,保质期一年。
Q:线粒体探针可以染植物、细菌吗?
A:该系列探针是针对动物细胞开发的,对于植物或是原核生物,由于存在细胞壁因而染色较为困难不是很方便,没有做过相关验证。
Q:线粒体探针染色后可以过夜在进行观察荧光吗?
A:该探针染色后应在 1h 内完成观察并拍照,过夜会导致荧光的消失,建议染色后立即进行观察以保证荧光拍照的结果。
Q:该线粒体探针染色后的固定剂是否可以更换为甲醇?
A:可以的。染色完成后,如需对细胞样本进行固定,除说明书建议外还可以使用预冷的100%甲醇于-20℃固定15min。
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