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Because cells were rinsed with PBS (or fresh cell culture medium) to remove extracellular compounds and contaminations, the detection of endogenous metabolites (e.g., cell lipids and adenosine phosphates) and exogenous compounds (e.g., anticancer drugs) [http://www.selleckchem.com/products/CP-673451.html see more] indicates that the Single-probe MS technique can be used to analyze intracellular compounds. Figure 1. Fabrication and setup of the single-probe for ambient MSI and SCMS analysis.a) Fabrication procedures of the single-probe. b) Photograph of a fabricated single-probe attached to a glass slide. c) Photograph of the single-probe setup attached to a mass spectrometer. d) Diagram of the single-probe setup coupled with a mass spectrometer. During an experiment, the sampling solvent is continuously provided from the syringe, the ionization voltage is applied to the conductive union from the mass spectrometer, two digital microscopes are used to monitor [http://www.selleckchem.com/products/azd5363.html click here] sample placement, the motorized XYZ stage system is used to control sample motion, and a mass spectrometer is used for analysis. e) Photograph of the customized digital stereoscope system. f) Photograph showing the digital stereoscope attached to the ion source interface flange through an optical board. Please click here to view a larger version of this figure. Figure 2. Results from an ambient MSI study of a mouse kidney section [https://en.wikipedia.org/wiki/Azastene Azastene] with high spatial and mass resolution.a) A representative mass spectrum from the single-probe MSI. The maximum intensity of detected metabolites can reach 3.39 x 107 (arbitrary units). b) A selection of the detected metabolites presented with their mass accuracy. c) MS images of [PC(32:0) + H]+ and [PC(34:1) + Na]+ taken from a mouse kidney section at 8.5 ?m spatial resolution. PC: phosphatidylcholine. Scale bar: 2 mm; 0.20 mm (inset). d) Determination of spatial resolution of the MS image for [PC(38:5) + Na]+ (adapted with permission from reference 15). Please click here to view a larger version of this figure. Figure 3. Results from an ambient SCMS analysis of drug treated HeLa cells with high mass resolution.a) Zoomed-in photograph of the single-probe tip showing a typical size of