Imaging of Cultured Cells by Mass Spectrometry

  • Hyun Jeong Yang
  • Yuki Sugiura
  • Koji Ikegami
  • Mitsutoshi Setou


Establishment of an Imaging mass spectrometry (IMS) experimental procedure for cultured cells is an important issue because it provides information on localization of various types of biomolecules inside the cell. At present, how to prepare the samples from cultured cells for an IMS has been under study. In this section, we present the preparation method for IMS analysis of mouse superior cervical ganglion (SCG) explant culture, containing many sympathetic neurons. The SCG explant culture is much larger than a single cell and neurons are highly polarized cells, allowing the comparison of the distinct compartment of the cells by IMS. The quick freeze-dry method was applied for the fixation of the neurons, and distinct distributions of small metabolites in the neurons were successfully visualized. We found that molecular composition was remarkably different between the cell body-containing region and the axon-enriched region.


Explant Culture Sympathetic Neuron Imaging Mass Spectrometry Superior Cervical Ganglion Matrix Application 
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  1. 1.
    Ostrowski SG, Van Bell CT, Winograd N, Ewing AG (2004) Mass spectrometric imaging of highly curved membranes during Tetrahymena mating. Science 305:71–73PubMedCrossRefGoogle Scholar
  2. 2.
    Rubakhin SS, Greenough WT, Sweedler JV (2003) Spatial profiling with MALDI MS: distribution of neuropeptides within single neurons. Anal Chem 75(20):5374–5380PubMedCrossRefGoogle Scholar
  3. 3.
    Kruse R, Sweedler JV (2003) Spatial profiling invertebrate ganglia using MALDI MS. J Am Soc Mass Spectrom 14(7):752–759PubMedCrossRefGoogle Scholar
  4. 4.
    Colliver TL, Brummel CL, Pacholski ML, et al. (1997) Atomic and molecular imaging at the single-cell level with TOF-SIMS. Anal Chem 69(13):2225–2231PubMedCrossRefGoogle Scholar
  5. 5.
    Ostrowski SG, Kurczy ME, Roddy TP, et al. (2007) Secondary ion MS imaging to relatively quantify cholesterol in the membranes of individual cells from differentially treated populations. Anal Chem 79(10):3554–3560PubMedCrossRefGoogle Scholar
  6. 6.
    Roddy TP, Cannon DM, Meserole CA, et al. (2002) Imaging of freeze-fractured cells with in situ fluorescence and time-of-flight secondary ion mass spectrometry. Anal Chem 74(16):4011–4019PubMedCrossRefGoogle Scholar
  7. 7.
    Roddy TP, Cannon DM, Ostrowsk SG, et al. (2002) Identification of cellular sections with imaging mass spectrometry following freeze fracture. Anal Chem 74(16):4020–4026PubMedCrossRefGoogle Scholar
  8. 8.
    Subhash C, George HM (1995) Imaging ion and molecular transport at subcellular resolution by secondary ion mass spectrometry. Int J Mass Spectrom Ion Process 143:161–176CrossRefGoogle Scholar
  9. 9.
    Liu Q, Guo Z, He L (2007) Mass spectrometry imaging of small molecules using desorption/ ionization on silicon. Anal Chem 79(10):3535–3541PubMedCrossRefGoogle Scholar
  10. 10.
    Chaurand P, Schriver KE, Caprioli RM (2007) Instrument design and characterization for high resolution MALDI-MS imaging of tissue sections. J Mass Spectrom 42(4):476–489PubMedCrossRefGoogle Scholar
  11. 11.
    Altelaar AFM, Taban IM, McDonnell LA, et al. (2007) High-resolution MALDI imaging mass spectrometry allows localization of peptide distributions at cellular length scales in pituitary tissue sections. Int J Mass Spectrom 260(2–3):203–211Google Scholar
  12. 12.
    Shimma S, Sugiura Y, Hayasaka T, et al. (2007) MALDI-based imaging mass spectrometry revealed abnormal distribution of phospholipids in colon cancer liver metastasis. J Chromatogr B 855(1):98–103CrossRefGoogle Scholar
  13. 13.
    Sugiura Y, Shimma S, Setou M (2006) Two-step matrix application technique to improve ionization efficiency for matrix-assisted laser desorption/ionization in imaging mass spec-trometry. Anal Chem 78(24):8227–8235PubMedCrossRefGoogle Scholar
  14. 14.
    Altelaar AFM, 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(3):734–742PubMedCrossRefGoogle Scholar
  15. 15.
    Hillebrandt H, Abdelghani A, Abdelghani-Jacquin C, et al. (2001) Electrical and optical characterization of thrombin-induced permeability of cultured endothelial cell monolayers on semiconductor electrode arrays. Appl Phys A Mater Sci Processing 73(5):539–546CrossRefGoogle Scholar
  16. 16.
    Qiu Q, Sayer M, Kawaja M, et al. (1998) Attachment, morphology, and protein expression of rat marrow stromal cells cultured on charged substrate surfaces. J Biomed Mater Res 42(1):117–127PubMedCrossRefGoogle Scholar
  17. 17.
    Aoki T, Tanino M, Sanui K, et al. (1996) Secretory function of adrenal chromaffin cells cultured on polypyrrole films. Biomaterials 17(20):1971–1974PubMedCrossRefGoogle Scholar
  18. 18.
    Yang H.J., Zaima N., Sugiura Y. et al. (in press), Imaging of cultured neurons by mass spectrometry., Proceedings for 7th International Symposium on Atomic Level Characteristions for New Materials and DevicesGoogle Scholar

Copyright information

© Springer 2010

Authors and Affiliations

  • Hyun Jeong Yang
    • 1
    • 2
  • Yuki Sugiura
    • 1
    • 2
  • Koji Ikegami
    • 2
  • Mitsutoshi Setou
    • 2
  1. 1.Department of Bioscience and BiotechnologyTokyo Institute of TechnologyMidori-ku, YokohamaJapan
  2. 2.Department of Molecular AnatomyHamamatsu University School of MedicineHigashi-ku, HamamatsuJapan

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