Journal Article

Correlated cryo-SEM and CryoNanoSIMS imaging of biological tissue

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

2023

Overview

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 methods all result in some degree of tissue morphology distortion and loss of soluble compounds. To overcome these limitations an entirely cryogenic sample preparation and imaging workflow is required. 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 15 N-enriched ammonium. 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.

• Publisher: Springer Science and Business Media LLC,BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Ammonium / ammonium assimilation / Automation / Biology (General) / Biomedical and Life Sciences / Chemical composition / Cooling / Cryoelectron Microscopy / Cryoelectron Microscopy; Microscopy, Electron, Scanning; High pressure freezing; Hydra viridissima; Isotope labeling; NanoSIMS; Osmoregulation; Photosymbiosis / elements / Freezing / Fresh water / green hydra / High pressure / High pressure freezing / Hydra viridissima / Image processing / Image resolution / ion mass-spectrometry / Isotope labeling / Isotopes / Life Sciences / Mapping / Mass spectrometry / Mass spectroscopy / Methodology / Methodology Article / Methods / Microscopy, Electron, Scanning / Morphology / NanoSIMS / Nitrogen / Nitrogen isotopes / Observations / Osmoregulation / Photosymbiosis / Planing / Polyvinyl alcohol / QH301-705.5 / Sample preparation / Scanning electron microscopy / Scanning microscopy / Secondary ion mass spectrometry / Tissues / Trace elements / Ultrastructure / Vitrification / Workflow