Time of flight mass spectrometry imaging of samples fractured in situ with a spring-loaded trap system.
Time of flight mass spectrometry imaging of samples fractured in situ with a spring-loaded trap system.
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DOI：
10.1021/ac101243b
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发表时间：
2010-08-01
影响因子：
7.4
通讯作者：
Ewing, Andrew G.
中科院分区：
文献类型：
--
作者：
Lanekoff, Ingela;Kurczy, Michael E.;Hill, Rowland;Fletcher, John S.;Vickerman, John C.;Winograd, Nick;Sjoevall, Peter;Ewing, Andrew G.
一种具有弹簧加载捕获系统的原位冷冻断裂装置已被设计出来，并用于单细胞的飞行时间二次离子质谱（TOF SIMS）分析。该装置采用夹层组件，这种组件通常用于冷冻断裂飞行时间二次离子质谱实验，以便为高分辨率二次离子质谱成像制备冷冻的水合细胞。在夹层组件中添加弹簧加载捕获系统为这种样品制备方法带来了两个改进。首先，通过添加弹簧使断裂机械化，无需使用低温冷却的刀具手动移除夹层组件的顶部，从而使每次断裂标准化。第二个改进是因为在夹层组件断裂后，夹层的顶部不会被丢弃。这就产生了两个成像表面，有效地使样品量翻倍，并提供了对因断裂而分叉的细胞的两个部分进行成像的独特能力。在此，我们报道了使用铋团簇离子源对冷冻断裂的大鼠嗜铬细胞瘤（PC12）细胞进行的飞行时间二次离子质谱分析。这项工作展示了从保存在冰基质中的细胞获得具有亚细胞横向分辨率的单细胞化学图像的能力。除了保存细胞外，在这些实验的冷冻断裂样品中，能更好地观察到在单细胞中很少被识别的脂质碎片离子的信号。此外，根据钾离子信号表明细胞已通过细胞质断裂这一公认的观点，我们还确定了细胞表面的不同断裂平面。将机械化的冷冻断裂装置与高分辨率团簇二次离子质谱成像相结合，将提供进行具有生物学相关性的二次离子质谱实验所需的灵敏度、分辨率以及试验次数。
An in situ freeze fracture device featuring a spring-loaded trap system has been designed and characterized for TOF SIMS analysis of single cells. The device employs the sandwich assembly, which is typically used in freeze fracture TOF SIMS experiments to prepare frozen, hydrated cells for high-resolution SIMS imaging. The addition of the spring-loaded trap system to the sandwich assembly offers two advances to this sample preparation method. First, mechanizing the fracture by adding a spring standardizes each fracture by removing the need to manually remove the top of the sandwich assembly with a cryogenically cooled knife. A second advance is brought about because the top of the sandwich is not discarded after the sandwich assembly has been fractured. This results in two imaging surfaces effectively doubling the sample size and providing the unique ability to image both sections of a cell bifurcated by the fracture. Here we report TOF SIMS analysis of freeze fractured rat pheochromocytoma (PC12) cells using a Bi cluster ion source. This work exhibits the ability to obtain single cell chemical images with sub-cellular lateral resolution from cells preserved in an ice matrix. In addition to preserving the cells, the signal from lipid fragment ions rarely identified in single cells are better observed in the freeze-fractured samples for these experiments. Furthermore, using the accepted argument that K+ signal indicates a cell that has been fractured though the cytoplasm, we have also identified different fracture planes of cells over the surface. Coupling a mechanized freeze fracture device to high-resolution cluster SIMS imaging will provide the sensitivity and resolution as well as the number of trials required to carry out biologically relevant SIMS experiments.
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影响因子：
2
作者：
Cliff, B;Lockyer, N;Vickerman, JC
通讯作者：
Vickerman, JC
影响因子：
7.4
作者：
Malm, Jakob;Giannaras, Dimitrios;Sjovall, Peter
通讯作者：
Sjovall, Peter
影响因子：
2.5
作者：
Nygren, Hakan;Hagenhoff, Birgit;Richter, Katrin
通讯作者：
Richter, Katrin
利用 TOF-SIMS 在单细胞水平上进行原子和分子成像
影响因子：
7.4
作者：
Colliver, TL;Brummel, CL;Winograd, N
通讯作者：
Winograd, N
影响因子：
7.4
作者：
Altelaar, AFM;Klinkert, I;Piersma, SR
通讯作者：
Piersma, SR