Abstract

The field of taxonomy is critically important for the identification, conservation, and ecology of biological species. Modern taxonomists increasingly need to employ advanced imaging techniques to classify organisms according to their observed morphological features. Moreover, the generation of three-dimensional datasets is of growing interest; moving beyond qualitative analysis to true quantitative classification. Unfortunately, biological samples are highly vulnerable to degradation under the energetic probes often used to generate these datasets. Neutral atom beam microscopes avoid such damage due to the gentle nature of their low energy probe, but to date have not been capable of producing three-dimensional data. Here we demonstrate a means to recover the height information for samples imaged in the scanning helium microscope (SHeM) via the process of stereophotogrammetry. The extended capabilities, namely sparse three-dimensional reconstructions of features, were showcased via taxonomic studies of both flora (Arabidopsis thaliana) and fauna (Heterodontus portusjacksoni). In concert with the delicate nature of neutral helium atom beam microscopy, the stereophotogrammetry technique provides the means to derive comprehensive taxonomical data without the risk of sample degradation due to the imaging process.

Highlights

  • Taxonomy, the science and practice of classification based on shared characteristics[1], is a crucial tool for the identification of species[2] as well as their conservation and ecology[3,4,5,6]

  • Morphology-based taxonomy often relies on microscopy techniques to identify features too small to be resolved by the human eye, and in recent years the field has moved towards incorporating three-dimensional imaging techniques to offer additional description methods for taxonomic applications[10,11,12]

  • Neutral helium microscopy is an emerging imaging technique, attractive for a wide range of delicate samples that could otherwise be damaged under the energetic probes of established microscopies

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Summary

Introduction

The science and practice of classification based on shared characteristics[1], is a crucial tool for the identification of species[2] as well as their conservation and ecology[3,4,5,6]. Milli-electron volt beam energies (orders of magnitude lower than comparable beams of electrons or photons), in tandem with the inherent properties of the helium atom – electrically neutral, chemically inert, and a lack of interaction with electric and magnetic fields - yields an ideal probe for delicate material systems. These same properties ensure unambiguous surface sensitivity (no penetration into the bulk) and no surface charging, removing the need for intrusive sample preparations. A detailed discussion of the experimental setup is given in the Materials and Methods section

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