Investigation of psoriasis skin tissue by label-free multi-modal imaging: a case study on a phototherapy-treated patient.

Background: Psoriasis is a systemic inflammatory disease characterized by epidermal proliferation in the skin. Altered lipid metabolism is considered to be a central factor in the psoriatic etiopathogenesis. Thus, it is necessary to visualize chemical specificity of the samples for better medical diagnosis and treatment. Here, we investigate its role in the development of psoriatic lesions, before and after ultraviolet phototherapy, in a case study. Methods: The distribution and morphology of different lipids and fibrous proteins in psoriatic (lesional) tissues were visualized by two complementary label-free imaging techniques: 1) non-linear microscopy (NLM), providing images of lipids/proteins throughout the skin layers at submicrometer resolution; and 2) mass spectrometry imaging (MSI), offering high chemical specificity and hence the detection of different lipid species in the epidermal and dermal regions. A conventional method of histological evaluation was performed on the tissues, with no direct comparison with NLM and MSI. Results: Psoriatic tissues had a higher lipid content, mainly in cholesterol, in both the epidermal and dermal regions, compared to healthy tissues. Moreover, the collagen and elastin fibers in the psoriatic tissues had a tendency to assemble as larger bundles, while healthy tissues showed smaller fibers more homogeneously spread. Although phototherapy significantly reduced the cholesterol content, it also increased the amounts of collagen in both lesional and non-lesional tissues. Conclusion: This study introduces NLM and MSI as two complementary techniques which are chemical specific and can be used to assess and visualize the distribution of lipids, collagen, and elastin in a non-invasive and label-free manner.

Imaging of vitamin D in psoriatic skin using time-of-flight secondary ion mass spectrometry (ToF-SIMS): A pilot case study.

Recently it has been recognized that there is a need of investigating in detail the vitamin D synthesis and metabolism directly in the skin with respect to its possible autocrine and paracrine actions. The potential effects the active metabolite of vitamin D exerts in pathological skin conditions like psoriasis needs to be clarified. Under ultraviolet B (UVB) irradiation skin can autonomously synthesize, activate and degrade vitamin D. In this pilot case study, we used for the first time Time-of-flight secondary ion mass spectrometry (ToF-SIMS) in the analysis of skin biopsies from a patient with psoriasis before and after UVB phototherapy. We were able to visualize vitamin D3 and its metabolites in the skin and subcutaneous tissue. At the same time information about their localization at subcellular level and morphology of the skin was received. This study proves that ToF-SIMS is a promising powerful tool to be used when investigating vitamin D´s role in dermatological diseases through skin biopsies.

Food-induced changes of lipids in rat neuronal tissue visualized by ToF-SIMS imaging.

Time of flight secondary ion mass spectrometry (ToF-SIMS) was used to image the lipid localization in brain tissue sections from rats fed specially processed cereals (SPC). An IonTof 5 instrument equipped with a Bi cluster ion gun was used to analyze the tissue sections. Data from 15 brain samples from control and cereal-fed rats were recorded and exported to principal components analysis (PCA). The data clearly show changes of certain lipids in the brain following cereal feeding. PCA score plots show a good separation in lipid distribution between the control and the SPC-fed group. The loadings plot reveal that the groups separated mainly due to changes in cholesterol, vitamin E and c18:2, c16:0 fatty acid distribution as well as some short chain monocarboxylic fatty acid compositions. These insights relate to the working mechanism of SPC as a dietary supplement. SPC is thought to activate antisecretory factor (AF), an endogenous protein with regulatory function for inflammation and fluid secretion. These data provide insights into lipid content in brain following SPC feeding and suggest a relation to activating AF.

Imaging TOF-SIMS of rat kidney prepared by high-pressure freezing.

Phosphocholine, potassium ions, and sodium ions were localized in rat kidney with imaging TOF-SIMS. Tissue preparation was performed with high-pressure freezing, freeze-fracturing and freeze-drying. The distribution of sodium ions was visualized by imaging the signal at m/z 23 of positively charged secondary ions, and the distribution of potassium ions was visualized by imaging the signal at m/z 39. Potassium was found localized within cells of the proximal tubulus epithelium and within cells of the glomeruli. High signals of sodium ions were seen in the interstitial tissue and also in epithelial cells of the collecting ducts and in glomeruli. The overlay image showed that the distribution of sodium ions and potassium ions were largely complementary with color mixing in glomeruli and in the interstitium surrounding proximal tubules. The ion distribution was further analyzed by correlation analysis. Phosphocholine-containing phospholipids were visualized by imaging the phosphocholine head group at m/z 184 of positively charged ions. The m/z 184 signal shows a ubiquitous distribution with a high intensity of phosphocholine in epithelial cells. Overlay image of m/z 184, m/z 39, and m/z 23 and multivariate analysis showed that the localization of high levels of phosphocholine colocalizes with high levels of potassium ions, as expected for an ion with intracellular localization.