TY - JOUR
T1 - Subcellular imaging of isotopically labeled carbon compounds in a biological sample by ion microprobe (NanoSIMS)
AU - Clode, Peta
AU - Stern, RA
AU - Marshall, A.T.
PY - 2007
Y1 - 2007
N2 - 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.
AB - 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.
U2 - 10.1002/jemt.20409
DO - 10.1002/jemt.20409
M3 - Article
C2 - 17279515
SN - 1059-910X
VL - 70
SP - 220
EP - 229
JO - Microscopy Research and Technique
JF - Microscopy Research and Technique
IS - 3
ER -