Mapping the cellular distribution of labelled molecules by SIMS microscopy.
We took advantage of one of the main possibilities of ion microscopy, ie isotopic analysis, to study the cellular distribution of molecules labelled either with carbon 14 or with stable isotopes of low natural abundance such as nitrogen 15 and deuterium. The surface of the sample is bombarded with an ion beam (O2+, Cs+ etc). Secondary ions emitted from the sample are filtered by a mass spectrometer and the distribution of the labelling isotope is recorded. In this way, we obtained images showing the characteristic distribution of 14C-thymidine and D-arginine in human fibroblasts, and of 15N-adenine in organotypic cultures of human breast cancer cells. The spatial resolution on the acquired images was close to 0.1 micron when using the UPS-ONERA ion microprobe. The sensitivity of the method for detecting carbon 14 is far greater than that of autoradiography and the technique is both fast and quantitative. On the other hand, the capacity of ion microscopy for studying the tissular distribution of molecules labelled with stable isotopes, opens the way for biological and pharmacological tracer studies of human diseases.
Hindie
E
E
Laboratoire de Biophysique, SC27 de l'Inserm, Créteil, France.
Coulomb
B
B
Beaupain
R
R
Galle
P
P
D000225:Adenine / Q000032:analysis; D000328:Adult; D001120:Arginine / Q000032:analysis; D001943:Breast Neoplasms / Q000737:chemistry / Q000648:ultrastructure; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D003903:Deuterium; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000648:ultrastructure; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D009587:Nitrogen Isotopes; D013936:Thymidine / Q000032:analysis; D014407:Tumor Cells, Cultured
D002250:Carbon Radioisotopes; D009587:Nitrogen Isotopes; D001120:Arginine; D003903:Deuterium; D000225:Adenine; D013936:Thymidine