Correlated cryo-SEM and CryoNanoSIMS imaging of biological tissue

生物组织的关联低温 SEM 和 CryoNanoSIMS 成像

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作者:Anders Meibom, Florent Plane, Tian Cheng, Gilles Grandjean, Olivier Haldimann, Stephane Escrig, Louise Jensen, Jean Daraspe, Antonio Mucciolo, Damien De Bellis, Nils Rädecker, Cristina Martin-Olmos, Christel Genoud, Arnaud Comment

Background

The development of nanoscale secondary ion mass spectrometry (NanoSIMS) has revolutionized the study of biological tissues by enabling, e.g., the visualization and quantification of metabolic processes at subcellular length scales. However, the associated sample preparation

Conclusion

With a cryo-workflow that includes vitrification by high pressure freezing, cryo-planing of the sample surface, and cryo-SEM imaging, the CryoNanoSIMS enables correlative ultrastructure and isotopic or elemental imaging of biological tissues in their most pristine post-mortem state. This opens new horizons in the study of fundamental processes at the tissue- and (sub)cellular level. Teaser: CryoNanoSIMS: subcellular mapping of chemical and isotopic compositions of biological tissues in their most pristine post-mortem state.

Results

Here, we report the development of a CryoNanoSIMS instrument that can perform isotope imaging of both positive and negative secondary ions from flat block-face surfaces of vitrified biological tissues with a mass- and image resolution comparable to that of a conventional NanoSIMS. This capability is illustrated with nitrogen isotope as well as trace element mapping of freshwater hydrozoan Green Hydra tissue following uptake of 15N-enriched ammonium.

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