Abstract
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.
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References
Belu, A. M., Graham, D. J., Castner D. G. (2003) Time-of-flight secondary ion mass spectrometry: techniques and applications for the characterization of biomaterial surfaces. Biomaterials, 24, 3635–3653.
Lockyer, N. P., Vickerman J. C. (2004) Progress in cellular analysis using ToF-SIMS. Appl Surf Sci, 231–232, 377–384.
Rubakhin, S. S., Jurchen, J. C., Monroe, E. B., Sweedler, J. V. (2005) Imaging mass spectrometry: fundamentals and applications to drug discovery. Drug Discov Today, 10, 823–837.
Guerquin-Kern, J.-L., Wu, T.-D., Quintana, C., Croisy, A. (2005) Progress in analytical imaging of the cell by dynamic secondary ion mass spectrometry (SIMS microscopy). Biochimica et Biophysica Acta, 1724, 228–238.
Chaurand, P., Cornett, D. S., Caprioli, R. M. (2006) Molecular imaging of thin mammalian tissue sections by mass spectrometry. Curr Opin Biotechnol, 17, 431–436.
Reyzer, M. L., Caprioli, R. M. (2007) MALDI-MS-based imaging of small molecules and proteins in tissues. Curr Opin Chem Biol, 11, 29–35.
Elowitz, M. B., Levine, A. J., Siggia, E. D., Swain, P. S. (2002) Stochastic gene expression in a single cell. Science, 297, 1183–1186.
Luxembourg, S. L., Mize, T. H., McDonnell, L. A., Heeren, R. M. A. (2004) High-spatial resolution mass spectrometric imaging of peptide and protein distributions on a surface. Anal Chem, 76, 5339–5344.
Jurchen, J. C., Rubakhin, S. S., Sweedler, J. V. (2005) MALDI-MS imaging of features smaller than the size of the laser beam. J Am Soc Mass Spectrom, 16, 1654–1659.
Rubakhin, S. S., Greenough, W. T., Sweedler, J. V. (2003) Spatial profiling with MALDI MS: distribution of neuropeptides within single neurons. Anal Chem, 75, 5374–5380.
Romer, W., Wu, T.-D., Duchambon, P., et al. (2006) Sub-cellular localisation of a 15N-labelled peptide vector using NanoSIMS imaging. Appl Surf Sci, 252, 6925–6930.
Audinot, J.-N., Guignard, C., Migeon, H.-N., Hoffmann, L. (2006) Study of the mechanism of diatom cell division by means of 29Si isotope tracing. Appl Surfe Sci, 252, 6813–6815.
McMahon, G., Glassner, B. J., Lechene, C. P. (2006) Quantitative imaging of cells with multi-isotope imaging mass spectrometry (MIMS) – nanoautography with stable isotope tracers. Appl Surf Sci, 252, 6895–6906.
Ostrowski, S. G., Kurczy, M. E., Roddy, T. P., Winograd, N., Ewing, A. G. (2007) Secondary ion MS imaging to relatively quantify cholesterol in the membranes of individual cells from differentially treated populations. Anal Chem, 79, 3554–3560.
Fletcher, J. S., Lockyer, N. P., Vaidyanathan, S., Vickerman, J. C. (2007) ToF-SIMS 3D biomolecular imaging of Xenopus laevis oocytes using buckminsterfullerene (C60) primary ions. Anal Chem, 79, 2199–2206.
Fletcher, J. S., Rabbani, S., Henderson, A., et al. (2008) A new dynamic in mass spectral imaging of single biological cells. Anal Chem, 80, 9058–9064.
Malmberg, P., Kriegeskotte, C., Arlinghaus, H. F., et al. (2008) Depth profiling of cells and tissues by using C60 + and SF5 + as sputter ions. Appl Surf Sci, 255, 926.
Kulp, K. S., Berman, E. S. F., Knize, M. G., et al. (2006) Chemical and biological differentiation of three human breast cancer cell types using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Anal Chem, 78, 3651–3658.
Liu, Q., Guo, Z., He, L. (2007) Mass spectrometry imaging of small molecules using desorption/ionization on silicon. Anal Chem, 79, 3535–3541.
Arlinghaus, H. F., Kriegeskotte, C., Fartmann, M., Wittig, A., Sauerwein, W., Lipinsky, D. (2006) Mass spectrometric characterization of elements and molecules in cell cultures and tissues. Appl Surf Sci, 252, 6941–6948.
Coello, Y., Gunaratne, T. C., Dantus, M. (2009) Atmospheric pressure femtosecond laser imaging mass spectrometry. Proceedings of SPIE, 7182, 71821 W.
Wells, W. A. (2005) The invention of freeze fracture EM and the determination of membrane structure. J Cell Biol, 168, 174–175.
Steere, R. L. (1957) Electron microscopy of structural detail in frozen biological specimens. J Biophys Biochem Cytol, 3, 45–60.
Chandra, S., Morrison, G. H. (1992) Sample preparation of animal tissues and cell cultures for secondary ion mass spectrometry (SIMS) microscopy. Biol Cell, 74, 31–42.
Cannon, D. M. J., Pacholski, M. L., Winograd, N., Ewing, A. G. (2000) Molecule specific imaging of freeze-fractured, frozen-hydrated model membrane systems using mass spectrometry. J Am Chem Soc, 122, 603–610.
Piehowski, P. D., Kurczy, M. E., Willingham, D., et al. (2008) Freeze-etching and vapor matrix deposition for ToF-SIMS imaging of single cells. Langmuir, 24, 7906–7911.
Nygren, H., Eriksson, C., Malmberg, P., et al. (2003) A cell preparation method allowing subellular localization of cholesterol and phosphocholine with imaging ToF-SIMS. Colloids Surf B Biointerfaces, 30, 87–92.
Parry, S., Winograd, N. (2005) High-resolution ToF-SIMS imaging of eukaryotic cells preserved in a trehalose matrix. Anal Chem, 77, 7950–7957.
Altelaar, A. F. M., Klinkert, I., Jalink, K., et al. (2006) Gold-enhanced biomolecular surface imaging of cells and tissue by SIMS and MALDI mass spectrometry. Anal Chem, 78, 734–742.
Berman, E. S. F., Fortson, S. L., Checchi, K. D., et al. (2008) Preparation of Single cells for imaging/profiling mass spectrometry. J Am Soc Mass Spectrom, 19, 1230–1236.
Acknowledgments
The authors gratefully acknowledge the assistance of Ligang Wu, Kyle D. Checchi, James S. Felton, Kuang Jen J. Wu, Cynthia B. Thomas, and Donald J. Sirbuly with various aspects of this research. This work was performed under the auspices of the US Department of Energy by the University of California, Lawrence Livermore National Laboratory under Contract No. W-7405-Eng-48 and supported, in part, by BCRP 10IB-0077, 11NB-0178, and LDRD 04-ERD-104 (LLNL internal funding).
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Berman, E.S., Fortson, S.L., Kulp, K.S. (2010). Preparation of Single Cells for Imaging Mass Spectrometry. In: Rubakhin, S., Sweedler, J. (eds) Mass Spectrometry Imaging. Methods in Molecular Biology, vol 656. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-746-4_15
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DOI: https://doi.org/10.1007/978-1-60761-746-4_15
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