Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 20, Pages 10702-10710Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b01256
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Funding
- National Research Fund Luxembourg (FNR) [C14/MS/8345352]
- C4HEALTH [INTER/MERA/14/9822270]
- US NIH [P41GM103412]
- US NIH
- NIH [RO1 NS027177I]
- NIH/NIBIB National Resource for Imaging Mass Spectrometry [NIH/NIBIB 5P41 EB001974]
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Correlative microscopy combining various imaging modalities offers powerful insights into obtaining 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 labeled tisue). 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 Laplacian 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.
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