ABSTRACT
Brain slice preparations cultured in vitro have long been used as a simplified model for studying brain development, electrophysiology, neurodegeneration and neuroprotection. In this paper an open fluidic system developed for improved long term culturing of organotypic brain slices is presented. The positive effect of continuous flow of growth medium, and thus stability of the glucose concentration and waste removal, is simulated and compared to the effect of stagnant medium that is most often used in tissue culturing. Furthermore, placement of the tissue slices in the developed device was studied by numerical simulations in order to optimize the nutrient distribution. The device was tested by culturing transverse hippocampal slices from 7 days old NMRI mice for a duration of 14 days. The slices were inspected visually and the slices cultured in the fluidic system appeared to have preserved their structure better than the control slices cultured using the standard interface method.
Subject(s)
Hippocampus/growth & development , Microfluidics/methods , Tissue Culture Techniques/methods , Animals , Mice , Microfluidics/instrumentationABSTRACT
Protein nanofibrils and nanotubes are now widely accepted as having potential for use in the field of bionanotechnology. For this to be a feasible alternative to existing technologies, there is a need for a commercially viable source. Previous work has identified amyloid fibrils formed from crude crystallin proteins as such a source, since these fibrils can be produced in large quantities at a low cost. Applications include use of fibrils as templates for the formation of nanowires or as biosensing scaffolds. There remains a number of practical considerations, such as stability and the ability to control their arrangement. In this study, crude crystallin amyloid fibrils are shown to be stable in a range of biological and clean room solvents, with the fibril presence confirmed by transmission electron microscopy and the thioflavin T fluorescent assay. The fibrils were also immobilised between microelectrodes using dielectrophoresis, which enabled the recording of I-V curves for small numbers of fibrils. This investigation showed the fibrils to have low conductivity, with current values in the range of 10(-10) A recorded. This low conductivity could be increased through modification, or alternately, the fibrils could be used unmodified for applications where they can act as templates or high surface area nanoscaffolds.
Subject(s)
Amyloid/chemistry , Crystallins/chemistry , Electrophoresis/methods , Nanostructures/chemistry , Amyloid/metabolism , Animals , Crystallins/metabolism , Electric Conductivity , Gadiformes , Lens, Crystalline/chemistry , Microscopy, Electron, Transmission , Protein Stability , SolubilityABSTRACT
In this article we present a generalized theoretical model for the continuous separation of particles using the pinched flow fractionation method. So far the theoretical models have not been able to predict the separation of particles without the use of correction factors. In this article we present a model which is capable of predicting the separation from first principles. Furthermore we comment on the importance of the incorporation of the finite height of the micro fluidic channels in the models describing the system behavior. We compare our model with the experiment obtained by Seki et al. (J. Takagi, M. Yamada, M. Yasuda and M. Seki, Lab Chip, 2005, 5, 778-784).
