We describe a new procedure for the parallel mapping of selected metals in histologically characterized tissue samples. Mapping is achieved via image registration of digital data obtained from two neighbouring cryosections by scanning the first as a histological sample and subjecting the second to laser ablation inductively coupled plasma mass spectrometry. This computer supported procedure enables determination of the distribution and content of metals of interest directly in the chosen histological zones and represents a substantial improvement over the standard approach, which determines these values in tissue homogenates or whole tissue sections. The potential of the described procedure was demonstrated in a pilot study that analysed Zn and Cu levels in successive development stages of pig melanoma tissue using MeLiM (Melanoma-bearing-Libechov-Minipig) model. We anticipate that the procedure could be useful for a complex understanding of the role that the spatial distribution of metals plays within tissues affected by pathological states including cancer.

Central European Institute of Technology Brno University of Technology Technicka 3058 10 CZ 616 00 Brno Czech Republic

Central European Institute of Technology Masaryk University Kamenice 5 CZ 625 00 Brno Czech Republic

Czech Institue of Informatics Robotics and Cybernetics CTU Prague Jugoslávských partyzánů 1580 3 CZ 160 00 Prague 6 Czech Republic

Department of Chemistry and Biochemistry Mendel University in Brno Zemedelska 1 CZ 613 00 Brno Czech Republic

Department of Chemistry Faculty of Science Masaryk University Kotlarska 2 CZ 611 37 Brno Czech Republic

Department of Cybernetics Faculty of Electrical Engineering Czech Technical University Technicka 2 CZ 166 27 Prague 6 Czech Republic

Department of Food Chemistry and Toxicology Berlin Institute of Technology Gustav Meyer Allee 25 D 133 55 Berlin Germany

Laboratory of Tumour Biology Institute of Animal Physiology and Genetics Academy of Sciences of the Czech Republic v v i Rumburska 89 CZ 277 21 Libechov Czech Republic

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Formigari A., Gregianin E. & Irato P. The effect of zinc and the role of p53 in copper-induced cellular stress responses. J Appl Toxicol 33, 527–536 (2013). PubMed

McCord M. C. & Aizenman E. The role of intracellular zinc release in aging, oxidative stress, and Alzheimer’s disease. Front Aging Neurosci 6, 1–16 (2014). PubMed PMC

Prasad A. S.. Zinc: role in immunity, oxidative stress and chronic inflammation. Curr Opin Clin Nutr Metab Care 12, 646–652 (2009). PubMed

Gumulec J. et al.. Serum and Tissue Zinc in Epithelial Malignancies: A Meta-Analysis. PLoS One 9, 1–11 (2014). PubMed PMC

Planska D., Burocziova M., Strnadel J. & Horak V. Immunohistochemical Analysis of Collagen IV and Laminin Expression in Spontaneous Melanoma Regression in the Melanoma-Bearing Libechov Minipig. Acta Histochem Cytochem 48, 15–26 (2015). PubMed PMC

Horak V., Fortyn K., Hruban V. & Klaudy J. Hereditary melanoblastoma in miniature pigs and its successful therapy by devitalization technique. Cell Mol Biol 45, 1119–1129 (1999). PubMed

Geffrotin C. et al.. Opposite regulation of tenascin-C and tenascin-X in MeLiM swine heritable cutaneous malignant melanoma. Biochim Biophys Acta-Gen Subj 1524, 196–202 (2000). PubMed

Borovansky J. et al.. Biochemical characterization of a new melanoma model the minipig MeLiM strain. Melanoma Res 13, 543–548 (2003). PubMed

Vincent-Naulleau S. et al.. Clinical and histopathological characterization of cutaneous melanomas in the melanoblastoma-bearing Libechov minipig model. Pigm Cell Res 17, 24–35 (2004). PubMed

Vaculovic T. et al.. Influence of laser ablation parameters on trueness of imaging. Appl Surf Sci 351, 296–302 (2015).

Becker J. S. et al.. Imaging of copper, zinc, and other elements in thin section of human brain samples (Hippocampus) by laser ablation inductively coupled plasma mass spectrometry. Anal Chem 77, 3208–3216 (2005). PubMed

Becker J. S. et al.. Bioimaging of metals by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Mass Spectrom Rev 29, 156–175 (2010). PubMed

Latkoczy C., Muller Y., Schmutz P. & Gunther D. Quantitative element mapping of Mg alloys by laser ablation ICP-MS and EPMA. Appl Surf Sci 252, 127–132 (2005).

Halicz L. & Gunther D. Quantitative analysis of silicates using LA-ICP-MS with liquid calibration. J Anal At Spectrom 19, 1539–1545 (2004).

van Elteren J. T., Tennent N. H. & Selih V. S.. Multi-element quantification of ancient/historic glasses by laser ablation inductively coupled plasma mass spectrometry using sum normalization calibration. Anal Chim Acta 644, 1–9 (2009). PubMed

Crain J. S. & Gallimore D. L.. Inductively coupled plasma-mass spectrometry of synthetic elements - TC-99. Appl Spectrosc 46, 547–549 (1992).

Frick D. A. & Gunther D. Fundamental studies on the ablation behaviour of carbon in LA-ICP-MS with respect to the suitability as internal standard. J Anal At Spectrom 27, 1294–1303 (2012).

Riesop D., Hirner A. V., Rusch P. & Bankfalvi A. Zinc distribution within breast cancer tissue: A possible marker for histological grading? J Cancer Res Clin Oncol 141, 1321–1331 (2015). PubMed PMC

Zitova B. & Flusser J. Image registration methods: a survey. Image Vis Comput 21, 977–1000 (2003).

Preparata F. P. & Shamos M. I.. Computational Geometry Springer-Verlag (2012).

Corvi M. & Nicchiotti G. Multiresolution image registration I E E E, Computer Soc Press (1995).

Pinheiro J. C. & Bates M. D.. Mixed-effects models in S and S-PLUS. Springer (2006).

Bates D., Machler M., Bolker B. M. & Walker S. C.. Fitting Linear Mixed-Effects Models Using lme4. J Stat Softw 67, 1–48 (2015).

Thai H. T. et al.. A comparison of bootstrap approaches for estimating uncertainty of parameters in linear mixed-effects models. Pharm Stat 12, 129–140 (2013). PubMed

Dunn O. J.. Multiple comparisons among means. J Am Stat Assoc 56, 52–64 (1961).

Fisher G. L., Spitler L. E., McNeill K. L. & Rosenblatt L. S.. Serum copper and zinc levels in melanoma patients. Cancer 47, 1838–1844 (1981). PubMed

Ros-Bullon M. R., Sanchez-Pedreno S. & Martinez-Liarte J. H.. Serum zinc levels are increased in melanoma patients. Melanoma Res 8, 273–277 (1998). PubMed

Gorodetsky R., Sheskin J. & Weinreb A. Iron, copper, and zinc concentrations in normal skin and in various nonmalignant and malignant lesions. Int J Dermatol 25, 440–445 (1986). PubMed

Manga P. et al.. A role for tyrosinase-related protein 1 in 4-tert-butyl phenol-induced toxicity in melanocytes - Implications for vitiligo. Am J Pathol 169, 1652–1662 (2006). PubMed PMC

Garcia-Borron J. C. & Sanchez M. C. O. Biosynthesis of Melanins Wiley-V C H Verlag Gmbh (2011).

Yamaguchi Y., Brenner M. & Hearing V. J.. The regulation of skin pigmentation. J Biol Chem 282, 27557–27561 (2007). PubMed

Zalewska M., Trefon J. & Milnerowicz H. The role of metallothionein interactions with other proteins. Proteomics 14, 1343–1356 (2014). PubMed

Weinlich G. et al.. Comparison of metallothionein-overexpression with sentinel lymph node biopsy as prognostic factors in melanoma. J Eur Acad Dermatol Venereol 21, 669–677 (2007). PubMed

Weinlich G. & Zelger B. Metallothionein overexpression, a highly significant prognostic factor in thin melanoma. Histopathology 51, 280–283 (2007). PubMed

McRae R., Bagchi P., Sumalekshmy S. & Fahrni C. J.. In Situ Imaging of Metals in Cells and Tissues. Chem Rev 109, 4780–4827 (2009). PubMed PMC

Determination of Renal Distribution of Zinc, Copper, Iron, and Platinum in Mouse Kidney Using LA-ICP-MS

Acute Increases in Intracellular Zinc Lead to an Increased Lysosomal and Mitochondrial Autophagy and Subsequent Cell Demise in Malignant Melanoma

Melanoma-Bearing Libechov Minipig (MeLiM): The Unique Swine Model of Hereditary Metastatic Melanoma

MALDI MSI of MeLiM melanoma: Searching for differences in protein profiles