Using SIMS and MIMS in biological materials: application to higher plants.
Analytical imaging by secondary ion mass spectrometry allows the precise cartography of elements and isotopes at subcellular level in biological samples. Multielemental coacquisition from the same sputtered zone gives new prospects in the study of biological matrix, where many elements coexist and move according to cellular metabolism. For plant studies, localizing and quantifying compartmentations of mineral nutrients is important to understanding their metabolism. The use of stable isotopes as analogous markers is an efficient strategy of labeling for transport and metabolic studies. As for all microanalytical techniques, the stabilization of biological material is essential for the use of secondary ion mass spectrometry/multi-isotope imaging mass spectrometry microprobes. The techniques chosen are explained: decontamination, cryoprocessing, sections in embedded material (Arabidopsis thaliana, Brassicae) and analysis. In the Notes section, we will comment on the need to adapt the protocol described to the variety and complexity of the biological materials. Other proven protocols established in other laboratories for other type of organisms or biological questions will also be introduced.
Grignon
Nicole
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Biochimie et Physiologie Moléculaire des Plantes, Centre National de la Recherche Scientifique, Institut National de la Recherche Agronomique, Ecole Nationale Supérieure Agronomique de Montpellier, France.
D017360:Arabidopsis / Q000378:metabolism / Q000648:ultrastructure; D015925:Cryopreservation; D008867:Microtomy; D018515:Plant Leaves / Q000378:metabolism / Q000648:ultrastructure; D018517:Plant Roots / Q000378:metabolism / Q000648:ultrastructure; D010944:Plants / Q000378:metabolism* / Q000648:ultrastructure; D016613:Plastic Embedding; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*