| 1. |
- Ahmad Tajudin, Asilah, et al.
(författare)
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Integrated acoustic immunoaffinity-capture (IAI) platform for detection of PSA from whole blood samples.
- 2013
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0189 .- 1473-0197. ; 13:9, s. 1790-1796
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Tidskriftsartikel (refereegranskat)abstract
- On-chip detection of low abundant protein biomarkers is of interest to enable point-of-care diagnostics. Using a simple form of integration, we have realized an integrated microfluidic platform for the detection of prostate specific antigen (PSA), directly in anti-coagulated whole blood. We combine acoustophoresis-based separation of plasma from undiluted whole blood with a miniaturized immunoassay system in a polymer manifold, demonstrating improved assay speed on our Integrated Acoustic Immunoaffinity-capture (IAI) platform. The IAI platform separates plasma from undiluted whole blood by means of acoustophoresis and provides cell free plasma of clinical quality at a rate of 10 uL/min for an online immunoaffinity-capture of PSA on a porous silicon antibody microarray. The whole blood input (hematocrit 38-40%) rate was 50 μl min(-1) giving a plasma volume fraction yield of ≈33%. PSA was immunoaffinity-captured directly from spiked female whole blood samples at clinically significant levels of 1.7-100 ng ml(-1) within 15 min and was subsequently detected via fluorescence readout, showing a linear response over the entire range with a coefficient of variation of 13%.
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| 2. |
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| 3. |
- Ainla, Alar, 1982, et al.
(författare)
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A multifunctional pipette
- 2012
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Ingår i: Lab on a Chip - Miniaturisation for Chemistry and Biology. - : Royal Society of Chemistry (RSC). - 1473-0189 .- 1473-0197. ; 12:7, s. 1255-1261
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Tidskriftsartikel (refereegranskat)abstract
- Microfluidics has emerged as a powerful laboratory toolbox for biologists, allowing manipulation and analysis of processes at a cellular and sub-cellular level, through utilization of microfabricated features at size-scales relevant to that of a single cell. In the majority of microfluidic devices, sample processing and analysis occur within closed microchannels, imposing restrictions on sample preparation and use. We present an optimized non-contact open-volume microfluidic tool to overcome these and other restrictions, through the use of a hydrodynamically confined microflow pipette, serving as a multifunctional solution handling and dispensing tool. The geometries of the tool have been optimised for use in optical microscopy, with integrated solution reservoirs to reduce reagent use, contamination risks and cleaning requirements. Device performance was characterised using both epifluorescence and total internal reflection fluorescence (TIRF) microscopy, resulting in similar to 200 ms and similar to 130 ms exchange times at similar to 100 nm and similar to 30 mm distances to the surface respectively.
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| 4. |
- Ainla, Alar, 1982, et al.
(författare)
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Single-cell electroporation using a multifunctional pipette
- 2012
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Ingår i: Lab on a Chip - Miniaturisation for Chemistry and Biology. - : Royal Society of Chemistry (RSC). - 1473-0189 .- 1473-0197. ; 12:22, s. 4605-4609
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Tidskriftsartikel (refereegranskat)abstract
- We present here a novel platform combination, using a multifunctional pipette to individually electroporate single-cells and to locally deliver an analyte, while in their culture environment. We demonstrate a method to fabricate low-resistance metallic electrodes into a PDMS pipette, followed by characterization of its effectiveness, benefits and limits in comparison with an external carbon microelectrode.
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| 5. |
- Andersson, Helene, et al.
(författare)
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Microfabrication and microfluidics for tissue engineering : state of the art and future opportunities
- 2004
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 4:2, s. 98-103
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Forskningsöversikt (refereegranskat)abstract
- An introductory overview of the use of microfluidic devices for tissue engineering is presented. After a brief description of the background of tissue engineering, different application areas of microfluidic devices are examined. Among these are methods for patterning cells, topographical control over cells and tissues, and bioreactors. Examples where microfluidic devices have been employed are presented such as basal lamina, vascular tissue, liver, bone, cartilage and neurons. It is concluded that until today, microfluidic devices have not been used extensively in tissue engineering. Major contributions are expected in two areas. The first is growth of complex tissue, where microfluidic structures ensure a steady blood supply, thereby circumventing the well-known problem of providing larger tissue structures with a continuous flow of oxygen and nutrition, and withdrawal of waste products. The second, and probably more important function of microfluidics, combined with micro/nanotechnology, lies in the development of in vitro physiological systems for studying fundamental biological phenomena.
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| 6. |
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| 7. |
- Andersson, Martin, et al.
(författare)
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A microscopy-compatible temperature regulation system for single-cell phenotype analysis - demonstrated by thermoresponse mapping of microalgae
- 2021
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry. - 1473-0197 .- 1473-0189. ; 21:9, s. 1694-1705
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Tidskriftsartikel (refereegranskat)abstract
- This work describes a programmable heat-stage compatible with in situ microscopy for the accurate provision of spatiotemporally defined temperatures to different microfluidic devices. The heat-stage comprises an array of integrated thin-film Joule heaters and resistance temperature detectors (RTDs). External programming of the heat-stage is provided by a custom software program connected to temperature controllers and heater–sensor pairs. Biologically relevant (20–40 °C) temperature profiles can be supplied to cells within microfluidic devices as spatial gradients (0.5–1.5 °C mm−1) or in a time-varying approach via e.g. step-wise or sinusoidally varying profiles with negligible temperature over-shoot. Demonstration of the device is achieved by exposing two strains of the coral symbiont Symbiodinium to different temperature profiles while monitoring their single-cell photophysiology via chlorophyll fluorometry. This revealed that photophysiological responses to temperature depended on the exposure duration, exposure magnitude and strain background. Moreover, thermal dose analysis suggested that cell acclimatisation occurs under longer temperature (6 h) exposures but not under shorter temperature exposures (15 min). As the thermal sensitivity of Symbiodinium mediates the thermal tolerance in corals, our versatile technology now provides unique possibilities to research this interdependency at single cell resolution. Our results also show the potential of this heat-stage for further applications in fields such as biotechnology and ecotoxicology.
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| 8. |
- Andersson Svahn, Helene, et al.
(författare)
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Single cells or large populations?
- 2007
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 7:5, s. 544-546
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Tidskriftsartikel (refereegranskat)
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| 9. |
- Antfolk, Maria, et al.
(författare)
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Focusing of sub-micrometer particles and bacteria enabled by two-dimensional acoustophoresis.
- 2014
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0189. ; 14:15, s. 2791-2799
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Tidskriftsartikel (refereegranskat)abstract
- Handling of sub-micrometer bioparticles such as bacteria are becoming increasingly important in the biomedical field and in environmental and food analysis. As a result, there is an increased need for less labor-intensive and time-consuming handling methods. Here, an acoustophoresis-based microfluidic chip that uses ultrasound to focus sub-micrometer particles and bacteria, is presented. The ability to focus sub-micrometer bioparticles in a standing one-dimensional acoustic wave is generally limited by the acoustic-streaming-induced drag force, which becomes increasingly significant the smaller the particles are. By using two-dimensional acoustic focusing, i.e. focusing of the sub-micrometer particles both horizontally and vertically in the cross section of a microchannel, the acoustic streaming velocity field can be altered to allow focusing. Here, the focusability of E. coli and polystyrene particles as small as 0.5 μm in diameter in microchannels of square or rectangular cross sections, is demonstrated. Numerical analysis was used to determine generic transverse particle trajectories in the channels, which revealed spiral-shaped trajectories of the sub-micrometer particles towards the center of the microchannel; this was also confirmed by experimental observations. The ability to focus and enrich bacteria and other sub-micrometer bioparticles using acoustophoresis opens the research field to new microbiological applications.
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| 10. |
- Antypas, H., et al.
(författare)
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A universal platform for selection and high-resolution phenotypic screening of bacterial mutants using the nanowell slide
- 2018
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Ingår i: Lab on a Chip. - : ROYAL SOC CHEMISTRY. - 1473-0197 .- 1473-0189. ; 18:12, s. 1767-1777
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Tidskriftsartikel (refereegranskat)abstract
- The Petri dish and microtiter plate are the golden standard for selection and screening of bacteria in microbiological research. To improve on the limited resolution and throughput of these methods, we developed a universal, user-friendly platform for selection and high-resolution phenotypic screening based on the nanowell slide. This miniaturized platform has an optimal ratio between throughput and assay complexity, holding 672 nanowells of 500 nl each. As monoclonality is essential in bacterial genetics, we used FACS to inoculate each nanowell with a single bacterium in 15 min. We further extended the protocol to select and sort only bacteria of interest from a mixed culture. We demonstrated this by isolating single transposon mutants generated by a custom-made transposon with dual selection for GFP fluorescence and kanamycin resistance. Optical compatibility of the nanowell slide enabled phenotypic screening of sorted mutants by spectrophotometric recording during incubation. By processing the absorbance data with our custom algorithm, a phenotypic screen for growth-associated mutations was performed. Alternatively, by processing fluorescence data, we detected metabolism-associated mutations, exemplified by a screen for -galactosidase activity. Besides spectrophotometry, optical compatibility enabled us to perform microscopic analysis directly in the nanowells to screen for mutants with altered morphologies. Despite the miniaturized format, easy transition from nano- to macroscale cultures allowed retrieval of bacterial mutants for downstream genetic analysis, demonstrated here by a cloning-free single-primer PCR protocol. Taken together, our FACS-linked nanowell slide replaces manual selection of mutants on agar plates, and enables combined selection and phenotypic screening in a one-step process. The versatility of the nanowell slide, and the modular workflow built on mainstream technologies, makes our universal platform widely applicable in microbiological research.
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