| 1. |
- Zieba, Agata, et al.
(author)
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Bright-Field Microscopy Visualization of Proteins and Protein Complexes by In Situ Proximity Ligation with Peroxidase Detection
- 2010
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In: Clinical Chemistry. - : Oxford University Press (OUP). - 0009-9147 .- 1530-8561. ; 56:1, s. 99-110
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Journal article (peer-reviewed)abstract
- BACKGROUND: The in situ proximity ligation assay (PLA) allows a protein or protein complex to be represented as an amplifiable DNA molecule. Recognition is mediated by proximity probes consisting of antibodies coupled with oligonucleotides. Upon dual binding of the proximity probes, the oligonucleotides direct the formation of a circular DNA molecule, which is then amplified by rolling-circle replication. The localized concatemeric product is then detected with fluorescent probes. The in situ PLA enables localized detection of individual native proteins or interacting protein pairs in fixed cells or tissue sections, thus providing an important tool for basic and clinical research. METHODS: We used horseradish peroxidase (HRP)conjugated oligonucleotides to couple in situ PLA with enzymatic visualization of the localized detection event. RESULTS: We demonstrate the detection of protein complexes, both in cells and in tissue sections, and show that we can quantify the complexes with image-analysis software specially developed for recognizing HRP signals in bright-field microscopy images. We show that fluorescence and HRP signals produce equivalent results, both ill cultured cells and in tissue samples. CONCLUSIONS: The combination of in situ PLA with bright-field detection and automated image analysis allows the signals present to be Counted in an automated fashion and thus provides a sensitive and specific method for quantification of proteins and protein complexes with bright-field microscopy. With this approach, in situ PLA can be used without the requirement for expensive fluorescence microscopes, thereby avoiding problems with nonspecific fluorescence while maintaining compatibility with conventional histologic staining.
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| 2. |
- Anderson, Dan, 1943, et al.
(author)
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Microwave breakdown in RF devices
- 2006
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In: 33rd European Physical Society Conference on Plasma Physics, Rome, June 19-23, 2006. ; , s. 429-
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Conference paper (peer-reviewed)
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| 4. |
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| 5. |
- Jordan, Ulf, 1974, et al.
(author)
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Microwave breakdown in slots
- 2004
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In: IEEE Trans. Plasma Science. ; 32, s. 2250-
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Journal article (peer-reviewed)
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| 6. |
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| 7. |
- Jordan, Ulf, 1974
(author)
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Microwave Breakdown: Physics and Applications
- 2005
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Doctoral thesis (other academic/artistic)abstract
- Theories for microwave breakdown in gases, with emphasis on Air, are presented in this thesis. Underlying physical processes are described together with models suitable for scientific research and technical development. Application of the models, both to microwave devices where the breakdown phenomenon limits normal operation and to devices where breakdown is an inherent feature of proper operation, is discussed. Paper A gives analytical and numerical models for breakdown in resonant structures involving inhomogeneous electric fields. The theoretical predictions are verified by experiments. Paper B introduces a model for breakdown in electric fields with stochastically varying amplitude. The stochastic model together with classical deterministic models for pulsed breakdown are used to assess microwave breakdown in multi-carrier and modulated fields. Paper C presents analytical and experimental results on the breakdown properties of narrow metallic slots, with special emphasis on parameters important for protection against high power microwave (HPM) radiation. Paper D examines in detail the concept of effective diffusion length for the determination of breakdown thresholds. The consequences of inhomogeneous electric fields and pressure dependence are covered, together with a discussion on determining the diffusion length from breakdown threshold data. Paper E gives analytical estimates for the nonlinear characteristics of high-Q resonant cavities during breakdown and discharge stages. The results can be used for predicting the behaviour of filter resonators and plasma processing devices. Paper F treats the problem of breakdown around a metal wedge, presenting conditions for localised breakdown not to occur close to the wedge. Conversely, the analysis also indicates when a more thorough analysis is needed to accurately determine the microwave breakdown threshold due to localisation of the breakdown close to the wedge.
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| 9. |
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| 10. |
- Jordan, Ulf, 1974, et al.
(author)
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On the effective diffusion length for microwave breakdown
- 2004
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In: Proc. 5th International Workshop on Multipactor, Corona and Passive Intermodulation, September 12 – 14, 2005, ESTEC, Noordwijk, The Netherlands In: Space RF Hardware (MULCOPIM' 2005). ; , s. 243-249
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Conference paper (peer-reviewed)
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