Berman Elena S F, Fortson Susan L, Kulp Kristen S
Los Gatos Research, Mountain View, CA, USA.
Methods Mol Biol. 2010;656:253-65. doi: 10.1007/978-1-60761-746-4_15.
Characterizing the molecular contents of individual cells is critical for understanding fundamental mechanisms of biological processes. Imaging mass spectrometry (IMS) of biological systems has been steadily gaining popularity for its ability to create precise chemical images of biological samples, thereby revealing new biological insights and improving understanding of disease. In order to acquire mass spectral images from single cells that contain relevant molecular information, samples must be prepared such that cell-culture components, especially salts, are eliminated from the cell surface and that the cell contents are accessible to the mass spectrometer. We have demonstrated a cellular preparation technique for IMS that preserves the basic morphology of cultured cells, allows mass spectrometric chemical profiling of cytosol, and removes the majority of the interfering species derived from the cellular growth medium. Using this protocol, we achieve high-quality, reproducible IMS images from three diverse cell types: MCF7 human breast cancer cells, Madin-Darby canine kidney (MDCK) cells, and NIH/3T3 mouse fibroblasts. This preparation method allows rapid and routine IMS analysis of cultured cells, making possible a wide variety of experiments to further scientific understanding of molecular processes within individual cells.
表征单个细胞的分子成分对于理解生物过程的基本机制至关重要。生物系统的成像质谱（IMS）因其能够创建生物样品的精确化学图像，从而揭示新的生物学见解并增进对疾病的理解而越来越受欢迎。为了从包含相关分子信息的单细胞中获取质谱图像，必须对样品进行制备，以便从细胞表面去除细胞培养成分，尤其是盐类，并且使细胞内容物能够被质谱仪检测到。我们已经展示了一种用于IMS的细胞制备技术，该技术保留了培养细胞的基本形态，允许对细胞质进行质谱化学分析，并去除了大部分来自细胞生长培养基的干扰物质。使用该方案，我们从三种不同的细胞类型中获得了高质量、可重复的IMS图像：MCF7人乳腺癌细胞、Madin-Darby犬肾（MDCK）细胞和NIH/3T3小鼠成纤维细胞。这种制备方法允许对培养细胞进行快速且常规的IMS分析，从而使各种各样的实验成为可能，以进一步科学地理解单个细胞内的分子过程。
