Correlative microscopy combining Secondary Ion Mass Spectrometry and Electron Microscopy: Comparison of intensity-hue-saturation and laplace pyramid methods for image fusion

Auteurs

F. Vollnhals, J.-N. Audinot, T. Wirtz, M. Mercier-Bonin, I. Fourquaux, B. Schroeppel, U. Kraushaar, V. Lev-Ram, M. H. Ellisman, and S. Eswara

Référence

Analytical Chemistry, vol. 89, no. 20, pp. 10702-10710, 2017

Description

Correlative microscopy combining various imaging modalities offers powerful insights to obtain a comprehensive understanding of physical, chemical and biological phenomena. In this article, we investigate two approaches for image-fusion in the context of combining the inherently lower-resolution chemical images obtained using Secondary Ion Mass Spectrometry (SIMS) with the high-resolution ultrastructural images obtained using Electron Microscopy (EM). We evaluate the image fusion methods with three different case-studies selected to broadly represent the typical samples in life science research: (i) histology (unlabeled tissue), (ii) nanotoxicology, and (iii) metabolism (isotopically labelled tissue). We show that the Intensity-Hue-Saturation Fusion method often applied for EM-sharpening can result in serious image artifacts, especially in cases where different contrast mechanisms interplay. Here, we introduce and demonstrate Laplace Pyramid Fusion as a powerful and more robust alternative method for image fusion. Both physical and technical aspects of correlative image overlay and image-fusion specific to SIMS-based correlative microscopy are discussed in detail alongside the advantages, limitations and the potential artifacts. Quantitative metrics to evaluate the results of image fusion are also discussed.

Lien

doi: 10.1021/acs.analchem.7b01256