| 1. |
- Eriksson, Therese, 1979-
(författare)
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Organelle movement in melanophores: Effects of Panax ginseng, ginsenosides and quercetin
- 2009
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Panax ginseng is a traditional herb that has been used for over 2000 years to promote health and longevity. Active components of ginseng include ginsenosides, polysaccharides, flavonoids, polyacetylenes, peptides, vitamins, phenols and enzymes, of which the ginsenosides are considered to be the major bioactive constituents. Although widely used, the exact mechanisms of ginseng and its compounds remain unclear. In this thesis we use melanophores from Xenopus laevis to investigate the effects of Panax ginseng extract G115 and its constituents on organelle transport and signalling. Due to coordinated bidirectional movement of their pigmented granules (melanosomes), in response to defined chemical signals, melanophores are capable of fast colour changes and provide a great model for the study of intracellular transport. The movement is regulated by alterations in cyclic adenosine 3’:5’-monophosphate (cAMP) concentration, where a high or low level induce anterograde (dispersion) or retrograde (aggregation) transport respectively, resulting in a dark or light cell. Here we demonstrate that Panax ginseng and its constituents ginsenoside Rc and Rd and flavonoid quercetin induce a concentration-dependent anterograde transport of melanosomes. The effect of ginseng is shown to be independent of cAMP changes and protein kinase A activation. Upon incubation of melanophores with a combination of Rc or Rd and quercetin, a synergistic increase in anterograde movement was seen, indicating cooperation between the ginsenoside and flavonoid parts of ginseng. Protein kinase C (PKC) inhibitor Myristoylated EGF-R Fragment 651-658 decreased the anterograde movement stimulated by ginseng and ginsenoside Rc and Rd. Moreover, ginseng, but not ginsenosides or quercetin, stimulated an activation of 44/42-mitogen activated protein kinase (MAPK), previously shown to be involved in both aggregation and dispersion of melanosomes. PKC-inhibition did not affect the MAPK-activation, suggesting a role for PKC in the ginseng- and ginsenoside-induced dispersion but not as an upstream activator of MAPK.
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| 2. |
- Iqbal, Zafar
(författare)
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Optical Sensing With an Ordinary Mobile Phone
- 2012
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- A major portion of the world’s population (≈ 80% of the total) lives in developing countries where lab instruments such as spectrophotometers are not widely available as their purchasing as well as maintenance is normally unaffordable. On the other hand, there are now around five billion mobile phone subscriptions worldwide and the current generation of standard mobile phones has several capabilities to perform user-defined analysis. This thesis contains work with respect to asses potentials and weaknesses of a standard mobile phone for use as a simplified spectrophotometric unit (as both the light source and detector) to perform analysis in the visible region (400-700 nm). A part of the work has been the development of the necessary software to be able to use an ordinary mobile phone to study diffuse and specular reflectance properties of the targeted samples using phone’s screen as controllable illumination source and the front view camera for simultaneous collection of spectral information.Papers I-III contain exploratory work performed to assess the potential of using the mobile phone as an optical sensor system. Papers IV and V present studies of more basic character of the interactions between the light from the phone screen and the sample, in particular for liquid samples.In paper I, tests with a virtual array of chemical indicators having areas with different colours were performed. Optimization of the alignment of the sample and the distance between the camera and the sample were carried out and the influence of ambient light was investigated. The lateral resolution of the images enables optical readout of sensor arrays as well as arrays for diagnostics.In paper II, the potential of using the technique for direct measurement of properties related to the quality of drinking water, food and beverages was investigated. Liquid samples were prepared in deionized water. Coloured compounds such as iron(III)chloride and humic acid were analyzed in the concentration range 0-10 mg/liter and were classified by their reflectance profiles with respect to the contamination type and its concentration. Colourless arsenic(III) was analyzed by its bleaching reaction with iodine/starch. An alternative arsenic detection method based on measurement of discolouration of iron containing sand was demonstrated.In paper III, it has been demonstrated that mobile phones can be used for qualitative analysis of foods and beverages, such as cold drinks, meat, vegetables and milk in terms of general food quality, safety and authenticity.In paper IV, the ability of the mobile phone system to measure absorption properties of liquid solutions is investigated. Different concentrations of colored solutions (reactive blue 2, Congo red and Metanil yellow) give rise to measurement data that are well described by the Beer-Lambert law. This is surprising since the measurement conditions were far from ideal, with a light source that was strongly polychromatic and an illumination that was not a collimated light beam with homogeneous light intensity. By analyzing red, green and blue light that was transmitted through the liquid a unique signature for classification and quantification was obtained. Also the repeatability and accuracy of the measurements were investigated and were surprisingly good for such a simple system. Analyses of reflectance properties of colored solid samples are also included and were more complex with results being dependent on the morphology and colorimetric properties of the different types of these samples.In paper V, it is found that different parts of the image data contain different information about liquid samples. While one part of the image gives information about the absorption properties as investigated in detail in paper IV, another part gives information about the refractive index of the sample. Measurements of samples with varying refractive index show trends expected from the Fresnel equations at zero incidence angle. Combined information from the two areas of the image offers new possibilities to classify samples.
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| 3. |
- Ulrich, Christian, 1976-
(författare)
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Electric Fields for Surface Design and Chemical Analysis
- 2008
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- This thesis deals with the use of electric fields for evaluation and control of chemical systems. An electric field can result in the flow of charge across an interface between a metal and a solution, by means of chemical reactions. This interplay between electricity and chemistry, i.e. electrochemistry, is a field of crucial importance both within research and industry. Applications based on electrochemical principles encompass such diverse areas as batteries and fuel cells, pH electrodes, and the glucose monitor used by people suffering from diabetes.A major part of the present work concerns the use of static electric fields in solutions containing a non-contacted metal surface. In such a setup it is possible to control the extent of electrochemical reactions at different positions on the metal. This allows the formation and evaluation of various types of gradients on electrodes, via indirectly induced electrochemical reactions. This approach is a new and simple way of forming for instance molecular gradients on conducting surfaces. These are very advantageous in biomimetic research, because a gradient contains a huge amount of discrete combinations of for example two molecules. The basis for the technique is the use of bipolar electrochemistry. Briefly, a surface can become a bipolar electrode (an electrode that acts as both anode and cathode) when the electric field in the solution exceeds a certain threshold value, thereby inducing redox reactions at both ends. In our experiments, the driving force for these reactions will vary along the electrode surface. Since the result of an electrochemical reaction can be the deposition or removal of material from an electrode, bipolar electrochemistry can be used to create gradients of that material on a surface. In order to gain a deeper understanding of these processes, the potential and current density distributions at bipolar electrodes were investigated with different methods. Especially the use of imaging techniques was important for the visualization and analysis of the gradients. Using this knowledge, the formation of more complex gradients was facilitated, and the results were further compared to simulations based on simple conductivity models. These simulations also provided us with means to predict the behavior of new and interesting setup geometries for pattering applications.The other major part is more application driven and deals with the use of alternating electric fields for chemical analysis, a technique known as electrochemical impedance spectroscopy (EIS). In this work, EIS has been applied for the analysis of engine oils and industrial cutting fluids. Emphasis was placed on practical aspects of the measurement procedure, and on the evaluation of the results using statistical methods. It was for example shown that it was possible to simultaneously determine the amount of different contaminants in low conducting solutions. Generally, EIS is used to measure the impedance of a solution or a solid, often as a function of the frequency of the alternating electric field. The impedance of a system is closely correlated to its complex dielectric constant, and EIS can therefor be used to examine many chemical and physical processes. It is further well suited for characterizing low conducting media with little or no redox-active species. The evaluation of impedance data is often a quite complex task, which is why we have made use of statistical methods that drastically reduce the effort and quickly reveal significant intrinsic parameters.
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| 4. |
- Preechaburana, Pakorn
(författare)
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Optical devices and methods for distributed lab-on-a-chip analyses
- 2013
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Lab-on-a-chip (LOC) technologies entail the miniaturization of analytical systems, and the reduction of required sample and reagent volumes. LOC devices offer compact alternatives to classical instrumentation while delivering comparable performance and disposable formats. These aspects make disposable LOC a clear candidate to support distributed chemical sensing applications; however, the need of accessory services and readout obstructs the materialization of pervasively distributed LOC solutions.In this thesis methods and devices to solve this problem are investigated. A distinctive aspect of this work is the pursuit of solutions based on disposable LOC elements specifically conceived to exploit ubiquitous infrastructure for readout and evaluation.Consumer electronic devices, such as cell phones are ubiquitous platforms with residual capabilities that can be used for chemical sensing, if properly interfaced. This work investigates elements and tools needed to empower cell phones as readers of disposable LOC devices and commercial disposable tests.Access to flexible fabrication of LOC devices at low cost is an important requisite for testing ideas and implementing customized solutions. A first contribution in this thesis is the development of a platform for mask less fabrication of 3D microstructures, which coexists on a routine fluorescence microscope. This microscope projection lithography system (MPLS) is capable of controlled 3D micro structuring, including cavities and cantilever geometries, and the sealing of monolithic micro cavities to glass substrates as well as the connection to large scale service areas. This fabrication platform and other fabrication methods were used along this thesis to provide disposable optical and fluidic components.Besides custom-made LOC solutions there are well-established commercial disposable devices, which are essentially compatible with decentralized diagnosis, except for the use of specialized readers that confine them to medical centers. The implementation of high dynamic range (HDR) imaging with standard cell phones, using the phone screen to control exposure, shows that sensitivity and resolution can be boosted to permit robust evaluation of this type of disposable tests, in decentralized scenarios.Solutions employing commercial tests, which have not been designed for cell phone evaluation, are typically suboptimal and the investigation of customized LOC components occupies a central role in this thesis. Accordingly, one important aspect to evaluate LOC devices in compact configurations is to be able to image the LOC at a close distance from the phone camera, a condition for which phones cameras are not able to focus.In addition, different phone brands and models have different optical specifications, and a practical refocusing solution should adapt to all of them. In this work an adaptive lens concept, complemented by phone time-lapse acquisition, which can be integrated in disposable LOCs, is demonstrated.The implementation of sensitive detection methods, such as surface plasmon resonance (SPR), which is compatible with label free protocols that simplify sample conditioning, is central to the materialization of ubiquitous LOCs readable with cell phones. In this thesis a disposable optical coupler, conditioning illumination taken from the phone screen, is used to create an angle resolved SPR signal from a LOC, which is read with the phone front camera. Tested performance is comparable with commercial compact SPR modules and detection of β2 microglobulin, which is an established marker for cancer, inflammatory disorders, and kidney disease, is within the diagnostics range for blood and urine.Finally, fluorescence detection within classical LOC devices is tailored to be detectable with consumer cameras. In this case a disposable optical coupler and fluidics is designed to condition laser illumination into total internal reflection excitation, while DSLR and phone cameras capture optically separated fluorescence. The system configuration supports a broad dynamic range and HDR imaging enables localized resolution boost at selected detection ranges. Detection of free fucose, a diagnostic marker for liver cirrhosis and several cancer forms, is shown feasible with a HDR implementation, as one last example of practical LOC detection schemes for decentralized scenarios.
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| 5. |
- Testorf, Martin, 1972-
(författare)
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Melanophores : cell biophysics and sensor applications
- 2001
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- This thesis deals with studies of melanophores and melanosomes by means of a physical approach. Melanophores are pigment cells that give the black colour to many vertebrates, e.g. fishes, frogs, and reptiles. Covering large areas of the skin, these cells are approximately 0.1 mm in diameter, and they contain black pigment granules called melanosomes.The geometry and the electric charge of isolated melanosomes were two physical properties that were studied. The electric charge was measured by electrophoresis and was found to be (-1.7 ± 0.2)·10-16 Coulomb in average. The geometry of melanosomes was measured using Scanning Force Microscopy, SFM, and resulted in an ellipsoidal shape with an average major diameter of 840 ± 20 nm.Under nervous and hormonal control, melanophores rearrange the intracellular melanosomes from a scattered distribution, called dispersed, to a state where all melanosomes are accumulated in the cell centre, called the aggregated state. In this way, melanophores change from black towards transparent. This gives an animal the ability to change not only between being pale or dark, but also between different colours by using melanophores to cover and uncover the colours of different types of pigment cells from underlying layers.The volume of melanosomes was measured with SFM. This study resulted in a difference of 18% when individual melanosomes from aggregated and dispersed melanophores were measured separately.Magnetic field exposure of melanophores has been reported to affect the aggregation. However, contradicting results are presented in the literature. To clarify the possible effect of magnetic fields on melanophores, experiments by aggregating fish melanophores under exposure to strong (8 and 14 Tesla), homogenous, static magnetic fields were carried out. Both the magnetic field-induced Lorentz force on the charged melanosomes and the reorientation of the cytoskeleton were considered as possible explanations of any effects. Whenfield experiments were compared to control experiments with zero field, no difference in aggregation levels were found. However, a more irregular speed of aggregation was seen in the 8 Tesla field than in the control experiments.A theoretical model was developed to explain switch-like responses in biological systems. A switch-like response to a graded stimulation was sometimes seen in the case of melanophores but was shown not to have a very large so-called Hill coefficient. The model is simple in its approach. It may be applied to general phenomena and is based on the assumption of a simultaneous desorption of an activator (agonist; substrate molecule; ... ) and an inactivator (antagonist, inhibitor; ... ) caused by a collision or interaction between two effector molecules (e.g. receptors or enzymes).Melanosomes are also found in the human body and have a remarkably capacity to bind other molecules. A well-established forensic application of this is to detect (illegal) drugs that have bound to melanosomes in hair shafts. So far this application is only qualitative. This thesis includes a characterisation of the binding of flunitrazepam to melanin. Flunitrazepam is the active substance of Rohypnol, which is a sedative that is illegal in several countries and sometimes called the "date-rape-drug".Melanophores are excellent model systems for studies of cellular phenomena. Moreover, melanophores are also interesting in sensing aspects. The change from black to transparent is a highly visible response to substances in their surroundings and has previously been the measurand in melanophore-based biosensors. The physical approach of these studies of melanophores also had the objective of evaluating new biosensor solutions.
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