| 1. |
- Barrett, Michael P., et al.
(författare)
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Microfluidics-based approaches to the isolation of African trypanosomes
- 2017
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Ingår i: Pathogens. - : MDPI AG. - 2076-0817. ; 6:4
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Forskningsöversikt (refereegranskat)abstract
- African trypanosomes are responsible for significant levels of disease in both humans and animals. The protozoan parasites are free-living flagellates, usually transmitted by arthropod vectors, including the tsetse fly. In the mammalian host they live in the bloodstream and, in the case of human-infectious species, later invade the central nervous system. Diagnosis of the disease requires the positive identification of parasites in the bloodstream. This can be particularly challenging where parasite numbers are low, as is often the case in peripheral blood. Enriching parasites from body fluids is an important part of the diagnostic pathway. As more is learned about the physicochemical properties of trypanosomes, this information can be exploited through use of different microfluidic-based approaches to isolate the parasites from blood or other fluids. Here, we discuss recent advances in the use of microfluidics to separate trypanosomes from blood and to isolate single trypanosomes for analyses including drug screening.
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| 2. |
- Beech, Jason P., et al.
(författare)
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Morphology-based sorting-blood cells and parasites
- 2010
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Ingår i: 14th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2010, MicroTAS 2010. - 9781618390622 ; 2, s. 1343-1345
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Konferensbidrag (refereegranskat)abstract
- Morphology represents a hitherto unexploited source of specificity in microfluidic particle separation and may serve as the basis for label-free particle fractionation. There is a wealth of morphological changes in blood cells due to a wide range of clinical conditions, diseases, medication and other factors. Also, blood-borne parasites differ in morphology from blood cells. We present the use of Deterministic Lateral Displacement to create a chip-based, label-free diagnostic tool, capable of harvesting some of the wealth of information locked away in red blood cell morphology. We also use the device to separate the parasites that cause sleeping sickness from blood.
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| 3. |
- Holm, Stefan H., et al.
(författare)
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A high-throughput deterministic lateral displacement device for rapid and sensitive field-diagnosis of sleeping sickness
- 2012
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Ingår i: Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012. - 9780979806452 ; , s. 530-532
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Konferensbidrag (refereegranskat)abstract
- We present a simple and rapid microfluidic device capable of extracting and concentrating the parasite causing the fatal disease sleeping sickness (SS) from blood. The device is based on deterministic lateral displacement (DLD) and constructed with a single inlet with flow induced by an ordinary syringe. The simplicity is crucial as the device is intended for use in the resource depraved areas where the disease is endemic. With only one inlet an intricate design with multiple depths has been utilized to create a cell free stream from the blood plasma into which the parasites are forced and subsequently collected in a detection region. In order to maximize the sample volume up to 10 device layers were stacked on top of each other which resulted in a throughput of ∼10 μL/min. This allowed for an approximate time per test of below 15 min.
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| 4. |
- Holm, Stefan H., et al.
(författare)
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Multiple depths in a deterministic lateral displacement device for field-diagnosis of sleeping-sickness
- 2011
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Ingår i: 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2011, MicroTAS 2011. - 9781618395955 ; 1, s. 527-529
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Konferensbidrag (refereegranskat)abstract
- We present a simple and inexpensive device capable of extracting and concentrating the parasite causing sleeping sickness from blood. The device is aimed at being used in rural resource depraved areas where the disease is endemic; therefore simplicity is of paramount importance. The device is based on deterministic lateral displacement with a single inlet and flow induced by a syringe. Through an intricate design with multiple depths, a cell free stream is created from the blood plasma into which the parasites are guided and subsequently collected in a dedicated reservoir for observation.
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| 5. |
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| 6. |
- Akbari, Elham, et al.
(författare)
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SEPARATION OF CLUSTERS OF GROUP A STREPTOCOCCI USING DETERMINISTIC LATERAL DISPLACEMENT
- 2021
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Ingår i: MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. - 9781733419031 ; , s. 1201-1202
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Konferensbidrag (refereegranskat)abstract
- Differences in morphologies of bacteria and bacteria clusters are known to influence their pathogenicity. However, it is difficult to separate cells and cell clusters based on morphology using standard cell biological methods, making studies of the underlying mechanisms difficult. Here we report a simple label-free method for the continuous separation of clusters of group A streptococci, based on cluster size and morphology, using Deterministic Lateral Displacement (DLD). In general, this opens up for the generation of cell populations with heterogenicity in cluster size and physical properties.
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| 7. |
- Akbari, Elham, et al.
(författare)
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SEPARATION OF SINGLETS AND CLUSTERS OF GROUP A STREPTOCOCCI USING DETERMINISTIC LATERAL DISPLACEMENT AND FILTER SONICATION
- 2022
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Ingår i: MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences. - 9781733419048 ; , s. 306-307
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Konferensbidrag (refereegranskat)abstract
- Differences in morphologies of bacteria and bacteria clusters are thought to contribute to their virulence and colonization. However, the conventional standard cell biological methods cannot separate bacteria and bacteria clusters based on their morphologies and sizes, making studies of the underlying mechanisms difficult. Here we report a simple label-free method for the continuous separation of singlets and clusters, of group A streptococci, based on their size and morphology, using Deterministic Lateral Displacement and filter-sonication. In general, this opens up for the generation of cell populations with heterogenicity in cluster size and physical properties.
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| 8. |
- Arellano-Caicedo, Carlos, et al.
(författare)
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Habitat complexity affects microbial growth in fractal maze
- 2023
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Ingår i: Current biology : CB. - : Elsevier BV. - 1879-0445 .- 0960-9822. ; 33:8, s. 4-1458
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Tidskriftsartikel (refereegranskat)abstract
- The great variety of earth's microorganisms and their functions are attributed to the heterogeneity of their habitats, but our understanding of the impact of this heterogeneity on microbes is limited at the microscale. In this study, we tested how a gradient of spatial habitat complexity in the form of fractal mazes influenced the growth, substrate degradation, and interactions of the bacterial strain Pseudomonas putida and the fungal strain Coprinopsis cinerea. These strains responded in opposite ways: complex habitats strongly reduced fungal growth but, in contrast, increased the abundance of bacteria. Fungal hyphae did not reach far into the mazes and forced bacteria to grow in deeper regions. Bacterial substrate degradation strongly increased with habitat complexity, even more than bacterial biomass, up to an optimal depth, while the most remote parts of the mazes showed both decreased biomass and substrate degradation. These results suggest an increase in enzymatic activity in confined spaces, where areas may experience enhanced microbial activity and resource use efficiency. Very remote spaces showing a slower turnover of substrates illustrate a mechanism which may contribute to the long-term storage of organic matter in soils. We demonstrate here that the sole effect of spatial microstructures affects microbial growth and substrate degradation, leading to differences in local microscale spatial availability. These differences might add up to considerable changes in nutrient cycling at the macroscale, such as contributing to soil organic carbon storage.
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| 9. |
- Arellano-Caicedo, Carlos, et al.
(författare)
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Habitat geometry in artificial microstructure affects bacterial and fungal growth, interactions, and substrate degradation
- 2021
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 4:1
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Tidskriftsartikel (refereegranskat)abstract
- Microhabitat conditions determine the magnitude and speed of microbial processes but have been challenging to investigate. In this study we used microfluidic devices to determine the effect of the spatial distortion of a pore space on fungal and bacterial growth, interactions, and substrate degradation. The devices contained channels differing in bending angles and order. Sharper angles reduced fungal and bacterial biomass, especially when angles were repeated in the same direction. Substrate degradation was only decreased by sharper angles when fungi and bacteria were grown together. Investigation at the cellular scale suggests that this was caused by fungal habitat modification, since hyphae branched in sharp and repeated turns, blocking the dispersal of bacteria and the substrate. Our results demonstrate how the geometry of microstructures can influence microbial activity. This can be transferable to soil pore spaces, where spatial occlusion and microbial feedback on microstructures is thought to explain organic matter stabilization.
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| 10. |
- Arellano-Caicedo, Carlos, et al.
(författare)
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Quantification of growth and nutrient consumption of bacterial and fungal cultures in microfluidic microhabitat models
- 2024
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Ingår i: STAR Protocols. - 2666-1667. ; 5:1
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Tidskriftsartikel (refereegranskat)abstract
- Understanding microbes in nature requires consideration of their microenvironment. Here, we present a protocol for quantifying biomass and nutrient degradation of bacterial and fungal cultures (Pseudomonas putida and Coprinopsis cinerea, respectively) in microfluidics. We describe steps for mask design and fabrication, master printing, polydimethylsiloxane chip fabrication, and chip inoculation and imaging using fluorescence microscopy. We include procedures for image analysis, plotting, and statistics. For complete details on the use and execution of this protocol, please refer to Arellano-Caicedo et al. (2023).1
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