Method for Macromolecular Colocalization Using Atomic Recombination in Dynamic SIMS

Localizing two or more components of assemblies in biological systems requires both continued development of fluorescence techniques and invention of entirely new techniques. Candidates for the latter include dynamic secondary ion mass spectrometry (D-SIMS). The latest generation of D-SIMS, the Came...

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Bibliographic Details
Published in:The journal of physical chemistry. B 2008-05, Vol.112 (17), p.5534-5546
Main Authors: Legent, G, Delaune, A, Norris, V, Delcorte, A, Gibouin, D, Lefebvre, F, Misevic, G, Thellier, M, Ripoll, C
Format: Article
Language:English
Subjects:
Carbon Isotopes
Crystallization
Engineering Sciences
Escherichia coli - chemistry
Escherichia coli - metabolism
Escherichia coli Proteins - biosynthesis
Escherichia coli Proteins - chemistry
Escherichia coli Proteins - isolation & purification
Glycine - chemistry
Ions
Life Sciences
Materials
Mathematics
Models, Biological
Nitrogen Isotopes
Spectrometry, Mass, Secondary Ion - methods
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Summary:Localizing two or more components of assemblies in biological systems requires both continued development of fluorescence techniques and invention of entirely new techniques. Candidates for the latter include dynamic secondary ion mass spectrometry (D-SIMS). The latest generation of D-SIMS, the Cameca NanoSIMS 50, permits the localization of specific, isotopically labeled molecules and macromolecules in sections of biological material with a resolution in the tens of nanometers and with a sensitivity approaching in principle that of a single protein. Here we use two different systems, crystals of glycine and mixtures of proteins, to show that the formation of recombinant CN secondary ions under Cs bombardment can be exploited to create a new colocalization technique. We show experimentally that the formation of the recombinant 13C15N secondary ion between 13C- and 15N-labeled macromolecules is indeed an indicator of the distance between the interacting macromolecules and on their shape. We build up a convolution model of the mixing-recombination process in D-SIMS that allows quantitative interpretations of the distance-dependent formation of the recombinant CN. Our results show that macromolecules can be colocalized if they are within 2 nm of one another. We discuss the potential advantages of this new technique for biological applications.
ISSN:1520-6106
1520-5207
DOI:10.1021/jp7100489