Subcellular imaging of isotopically labeled carbon compounds in a biological sample by ion microprobe (NanoSIMS)

被引:49
作者
Clode, Peta L.
Stern, Richard A.
Marshall, Alan T.
机构
[1] Univ Western Australia, Ctr Microscopy & Microanal, Crawley, WA 6009, Australia
[2] Geosci Australia, GA Geochronol Lab, Minerals Div, Canberra, ACT, Australia
[3] La Trobe Univ, Analyt Electron Microscopy Lab, Melbourne, Vic, Australia
关键词
secondary ion mass spectrometry; molecular transport; isotopic tracers; ion imaging;
D O I
10.1002/jemt.20409
中图分类号
R602 [外科病理学、解剖学]; R32 [人体形态学];
学科分类号
100101 ;
摘要
Here we demonstrate the technique of nanoscale secondary ion mass spectrometry, utilizing the Cameca NanoSIMS50(R) ion microprobe, to detect and image the metabolism of an isotopically labeled compound ((NaHCO3)-C-13) in a biological sample. In particular, we have designed and verified protocols for imaging the subcellular distribution and determining the relative abundance of labeled C-13, within the coral Galaxea fascicularis. Analyses were conducted on 1-mu m thick sections of resin-embedded material, using both scanned (mapping) and static (spot analysis) Cs+ primary ion beam of similar to 100 nm diameter. Using these samples we establish that NanoSIMS has adequate mass resolution to reliably distinguish C-13 from potential isobaric interference by (CH)-C-12-H-1 and that data extracted from ion maps are comparable to those acquired by spot analyses. Independent of the method of acquisition, ratioing of C-13 to the naturally abundant C-12 is essential if meaningful data, which can be statistically compared to standard and control samples, are to be obtained. These results highlight the potential of NanoSIMS for intracellular tracking of a variety of organic and inorganic compounds labeled with stable isotopes of C, N, O, S, P, and halogens.
引用
收藏
页码:220 / 229
页数:10
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