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Pseudo-HE images derived from CARS/TPEF/SHG multimodal imaging in combination with Raman-spectroscopy as a pathological screening tool

2016, Bocklitz, Thomas W., Salah, Firas Subhi, Vogler, Nadine, Heuke, Sandro, Chernavskaia, Olga, Schmidt, Carsten, Waldner, Maximilian J., Greten, Florian R., Bräuer, Rolf, Schmitt, Michael, Stallmach, Andreas, Petersen, Iver, Popp, Jürgen

Due to the steadily increasing number of cancer patients worldwide the early diagnosis and treatment of cancer is a major field of research. The diagnosis of cancer is mostly performed by an experienced pathologist via the visual inspection of histo-pathological stained tissue sections. To save valuable time, low quality cryosections are frequently analyzed with diagnostic accuracies that are below those of high quality embedded tissue sections. Thus, alternative means have to be found that enable for fast and accurate diagnosis as the basis of following clinical decision making.

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Multimodal Nonlinear Microscopy for Therapy Monitoring of Cold Atmospheric Plasma Treatment

2019, Meyer, Tobias, Bae, Hyeonsoo, Hasse, Sybille, Winter, Jörn, von Woedtke, Thomas, Schmitt, Michael, Weltmann, Klaus-Dieter, Popp, Jürgen

Here we report on a non-linear spectroscopic method for visualization of cold atmospheric plasma (CAP)-induced changes in tissue for reaching a new quality level of CAP application in medicine via online monitoring of wound or cancer treatment. A combination of coherent anti-Stokes Raman scattering (CARS), two-photon fluorescence lifetime imaging (2P-FLIM) and second harmonic generation (SHG) microscopy has been used for non-invasive and label-free detection of CAP-induced changes on human skin and mucosa samples. By correlation with histochemical staining, the observed local increase in fluorescence could be assigned to melanin. CARS and SHG prove the integrity of the tissue structure, visualize tissue morphology and composition. The influence of plasma effects by variation of plasma parameters e.g., duration of treatment, gas composition and plasma source has been evaluated. Overall quantitative spectroscopic markers could be identified for a direct monitoring of CAP-treated tissue areas, which is very important for translating CAPs into clinical routine.

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A rigid coherent anti-Stokes Raman scattering endoscope with high resolution and a large field of view

2018, Zirak, P., Matz, Gregor, Messerschmidt, Bernhard, Meyer, Tobias, Schmitt, Michael, Popp, Jürgen, Uckermann, Ortrud, Galli, R., Kirsch, Matthias, Winterhalder, M.J., Zumbusch, A.

Nonlinear optical endoscopy is an attractive technique for biomedical imaging since it promises to give access to high resolution imaging in vivo. Among the various techniques used for endoscopic contrast generation, coherent anti-Stokes Raman scattering (CARS) is especially interesting. CARS endoscopy allows molecule specific imaging of unlabeled samples. In this contribution, we describe the design, implementation, and experimental characterization of a rigid, compact CARS endoscope with a spatial resolution of 750 nm over a field of view of roughly 250 μm. Omission of the relay optics and use of a gradient index lens specifically designed for this application allow one to realize these specifications in an endoscopic unit which is 2.2 mm wide over a length of 187 mm, making clinical applications during surgical interventions possible. Multimodal use of the endoscope is demonstrated with images of samples with neurosurgical relevance.Nonlinear optical endoscopy is an attractive technique for biomedical imaging since it promises to give access to high resolution imaging in vivo. Among the various techniques used for endoscopic contrast generation, coherent anti-Stokes Raman scattering (CARS) is especially interesting. CARS endoscopy allows molecule specific imaging of unlabeled samples. In this contribution, we describe the design, implementation, and experimental characterization of a rigid, compact CARS endoscope with a spatial resolution of 750 nm over a field of view of roughly 250 μm. Omission of the relay optics and use of a gradient index lens specifically designed for this application allow one to realize these specifications in an endoscopic unit which is 2.2 mm wide over a length of 187 mm, making clinical applications during surgical interventions possible. Multimodal use of the endoscope is demonstrated with images of samples with neurosurgical relevance.

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Shape-Memory Metallopolymer Networks Based on a Triazole–Pyridine Ligand

2019, Meurer, Josefine, Hniopek, Julian, Zechel, Stefan, Enke, Marcel, Vitz, Jürgen, Schmitt, Michael, Popp, Jürgen, Hager, Martin D., Schubert, Ulrich S.

Shape memory polymers represent an interesting class of stimuli-responsive polymers. With their ability to memorize and recover their original shape, they could be useful in almost every area of our daily life. We herein present the synthesis of shape-memory metallopolymers in which the switching unit is designed by using bis(pyridine–triazole) metal complexes. The polymer networks were synthesized via free radical polymerization of methyl-, ethyl- or butyl-methacrylate, tri(ethylene glycol) dimethacrylate and a methacrylate moiety of the triazole–pyridine ligand. By the addition of zinc(II) or cobalt(II) acetate it was possible to achieve metallopolymer networks featuring shape-memory abilities. The successful formation of the metal-ligand complex was proven by Fourier transform infrared (FT-IR) spectroscopy and by 1H NMR spectroscopy. Furthermore, the shape-recovery behavior was studied in detailed fashion and even triple-shape memory behavior could be revealed.

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Comparison of hyperspectral coherent Raman scattering microscopies for biomedical applications

2018, Bocklitz, Thomas W., Meyer, Tobias, Schmitt, Michael, Rimke, Ingo, Hoffmann, Franziska, von Eggeling, Ferdinand, Ernst, G., Guntinas-Lichius, Orlando, Popp, Jürgen

Raman scattering based imaging represents a very powerful optical tool for biomedical diagnostics. Different Raman signatures obtained by distinct tissue structures and disease induced changes provoke sophisticated analysis of the hyperspectral Raman datasets. While the analysis of linear Raman spectroscopic tissue data is quite established, the evaluation of hyperspectral nonlinear Raman data has not yet been evaluated in great detail. The two most common nonlinear Raman methods are CARS (coherent anti-Stokes Raman scattering) and SRS (stimulated Raman scattering) spectroscopy. Specifically the linear concentration dependence of SRS as compared to the quadratic dependence of CARS has fostered the application of SRS tissue imaging. Here, we applied spectral processing to hyperspectral SRS and CARS data for tissue characterization. We could demonstrate for the first time that similar cluster distributions can be obtained for multispectral CARS and SRS data but that clustering is based on different spectral features due to interference effects in CARS and the different concentration dependence of CARS and SRS. It is shown that a direct combination of CARS and SRS data does not improve the clustering results.