Chemical analysis of osmium tetroxide staining in adipose tissue using imaging ToF-SIMS |
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Authors: | Dalila Belazi Santiago Solé-Domènech Björn Johansson Martin Schalling Peter Sjövall |
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Affiliation: | 1.Department of Molecular Medicine and Surgery, Karolinska Institutet,Karolinska Hospital CMM L8:00,Stockholm,Sweden;2.Department of Chemistry and Materials Technology,SP Technical Research Institute of Sweden,Bor?s,Sweden |
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Abstract: | Osmium tetroxide (OsO4) is a commonly used stain for unsaturated lipids in electron and optical microscopy of cells and tissues. In this work, the localization of osmium oxide and specific lipids was independently monitored in mouse adipose tissue by using time-of-flight secondary ion mass spectrometry with Bi cluster primary ions. Substance-specific ion images recorded after OsO4 staining showed that unsaturated C18 fatty acids were colocalized with osmium oxide, corroborating the view that osmium tetroxide binds to unsaturated lipids. In contrast, saturated fatty acids (C14, C16 and C18) and also unsaturated C16 fatty acids show largely complementary localizations to osmium oxide. Furthermore, the distributions of saturated and unsaturated diglycerides are consistent with the specific binding of osmium oxide to unsaturated C18 fatty acids. The abundance of ions, characteristic of phospholipids and proteins, is strongly decreased as a result of the osmium staining, suggesting that a large fraction of these compounds are removed from the tissue during this step, while ions related to fatty acids, di- and triglycerides remain strong after osmium staining. Ethanol dehydration after osmium staining results in more homogeneous distributions of osmium oxide and unsaturated lipids. This work provides detailed insight into the specific binding of osmium oxide to different lipids. |
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Keywords: | ToF-SIMS Saturated fatty acids Unsaturated fatty acids Glycerides Osmium tetroxide |
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