Summary
3D chemical microscopy is one of the emerging applications of secondary ion mass spectrometry (SIMS) in biology. Tissues, cells, extracellular matrices, and polymer films can be imaged at present with a lateral resolution of 50 nm and depth resolution of 1 nm using the latest generation of CAMECA magnetic sector NanoSIMS 50 or with a lower lateral resolution (above 100 nm) using IMS 4f Cameca SIMS equipped with cold stage. Dynamic mode SIMS analysis is performed in ultrahigh vacuum and thus requires specific and careful preparation of biological samples aimed at preserving and minimizing destruction of the original structural and chemical properties of the samples. Here we describe a methodology based on the ultrafast plunge-freezing of biological tissues, preparation of the sample for SIMS analyses and transfer to the SIMS cold stage without interruption of the cold chain during the mounting procedure and subsequent SIMS analyses. Using this strategy, SIMS chemical microscopy can be performed on biological tissue in which unwanted molecular and/or structural reorganization, loss of constituents and chemical modifications are minimized and in which structures are therefore optimally preserved.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Dérue, C., Giboun, D., Demarty, M., Verdus, M.C., Lefebvre, F., Thellier, M., and Ripoll, C.(2006)Dynamic-SIMS imaging and quantification of inorganic ions in frozen-hydrated plant samples. Microscopy Research and Techniques69, 53–63.
Dérue, C., Giboun, D., Verdus, M.C., Lefebvre, F., Demarty, M., Ripoll, C., and Thellier, M.(2002)Appraisal of SIMS applicability to boron studies in plants. Microscopy Research and Techniques58, 104–110.
Dérue, C., Gibouin, D., Lefebvre, F., Rasser, B., Robin, A., Le Sceller, L., Verdus, M.-C., Demarty, M., Thellier, M., and Ripoll, C.(1999)A new cold stage for SIMS analysis and imaging of frozen-hydrated biological samples. Journal of Trace and Microprobe Techniques17, 451–460.
Dérue, C., Gibouin, D., Lefebvre, F., Studer, D., Thellier, M., and Ripoll, C.(2006)Relative sensitivity factors of inorganic cations in frozen-hydrated standards in secondary ion MS analysis. Analytical Chemistry78, 2471–2477.
Acknowledgments
This work was supported by the Région de Haute Normandie, the Ministère de la Recherche et de la Technologie, the CNRS, the University of Rouen, and the sixth framework programme Network of Excellence Nanobeams.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Misevic, G. et al. (2009). Chemical Microscopy of Biological Samples by Dynamic Mode Secondary Ion Mass Spectrometry (SIMS). In: Even-Ram, S., Artym, V. (eds) Extracellular Matrix Protocols. Methods in Molecular Biology, vol 522. Humana Press. https://doi.org/10.1007/978-1-59745-413-1_10
Download citation
DOI: https://doi.org/10.1007/978-1-59745-413-1_10
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-984-0
Online ISBN: 978-1-59745-413-1
eBook Packages: Springer Protocols