| 1. |
- Afrasiabi, Roodabeh, et al.
(författare)
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Integration of a droplet-based microfluidic system and silicon nanoribbon FET sensor
- 2016
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Ingår i: Micromachines. - : MDPI AG. - 2072-666X. ; 7:8
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Tidskriftsartikel (refereegranskat)abstract
- We present a novel microfluidic system that integrates droplet microfluidics with a silicon nanoribbon field-effect transistor (SiNR FET), and utilize this integrated system to sense differences in pH. The device allows for selective droplet transfer to a continuous water phase, actuated by dielectrophoresis, and subsequent detection of the pH level in the retrieved droplets by SiNR FETs on an electrical sensor chip. The integrated microfluidic system demonstrates a label-free detection method for droplet microfluidics, presenting an alternative to optical fluorescence detection. In this work, we were able to differentiate between droplet trains of one pH-unit difference. The pH-based detection method in our integrated system has the potential to be utilized in the detection of biochemical reactions that induce a pH-shift in the droplets.
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| 2. |
- Björk, Sara, 1990-, et al.
(författare)
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Droplet microfluidic microcolony sorting by fluorescence area for high throughput, yield-based screening of triacyl glycerides in S. Cerevisiae
- 2020
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Ingår i: MicroTAS 2020 - 24th International Conference on Miniaturized Systems for Chemistry and Life Sciences. - : Chemical and Biological Microsystems Society. ; , s. 1015-1016
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Konferensbidrag (refereegranskat)abstract
- Here we present a droplet microfluidics workflow for cell factory screening by yield of an intracellular product from isogenic microcolonies, i.e. minimal cell populations, encapsulated in picoliter droplets. This allows us to utilize all the benefits of droplet microfluidic screening in terms of throughput, but based on the signal from a population average, rather than the noisy single cell signal. We demonstrate microcolony sorting by integrated droplet fluorescence area of encapsulated E. coli, optimize triglyceride (TG) microcolony assay in droplets and apply the microcolony screening concept to analyze triglyceride (TG) production in S. cerevisiae.
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| 3. |
- Damiati, Safa A., et al.
(författare)
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Artificial intelligence application for rapid fabrication of size-tunable PLGA microparticles in microfluidics
- 2020
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Ingår i: Scientific Reports. - : Springer Nature. - 2045-2322. ; 10:1
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Tidskriftsartikel (refereegranskat)abstract
- In this study, synthetic polymeric particles were effectively fabricated by combining modern technologies of artificial intelligence (AI) and microfluidics. Because size uniformity is a key factor that significantly influences the stability of polymeric particles, therefore, this work aimed to establish a new AI application using machine learning technology for prediction of the size of poly(d,l-lactide-co-glycolide) (PLGA) microparticles produced by diverse microfluidic systems either in the form of single or multiple particles. Experimentally, the most effective factors for tuning droplet/particle sizes are PLGA concentrations and the flow rates of dispersed and aqueous phases in microfluidics. These factors were utilized to develop five different and simple in structure artificial neural network (ANN) models that are capable of predicting PLGA particle sizes produced by different microfluidic systems either individually or jointly merged. The systematic development of ANN models allowed ultimate construction of a single in silico model which consists of data for three different microfluidic systems. This ANN model eventually allowed rapid prediction of particle sizes produced using various microfluidic systems. This AI application offers a new platform for further rapid and economical exploration of polymer particles production in defined sizes for various applications including biomimetic studies, biomedicine, and pharmaceutics.
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| 4. |
- Höjer, Pontus, et al.
(författare)
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Identification of Major Immune Cell Lineages with DBS-Pro
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Proteins play a pivotal role in cellular function and heterogeneity. Understanding cellular diversity at the proteome level necessitates sensitive single-cell assays with high throughput. While current sequencing-based methods offer promise, they often face limitations, including reliance on expensive and inaccessible commercial platforms. Here, we have adopted the DBS-Pro method, utilizing site-specific oligonucleotide-conjugated antibodies, to analyze surface proteins in single cells. The method uses cheap degenerated barcode oligonucleotides and a simple microfluidics setup for cell encapsulation. A sample of PBMCs was examined using a panel targeting six separate immune cell markers. Using this panel we could quantify marker expression on 1,307 cells, identifying major immune cell lineages including CD4+ T-cells, CD8+ T-cells, monocytes, and B-cells. While recognizing the need for protocol improvements, our results present a promising approach for single-cell proteomics.
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| 5. |
- Langer, Krzysztof, et al.
(författare)
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A conversational robotic lab assistant for automated microfluidic 3d microtissue production
- 2019
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Ingår i: 23rd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2019. - : Chemical and Biological Microsystems Society. ; , s. 888-889
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Konferensbidrag (refereegranskat)abstract
- The future of life science is linked to automation and microfluidics. Here we present a robotic lab assistant, a general automation platform for droplet microfluidics including a conversational mobile interface. We demonstrate the automated production of human cancer microtissues in droplets at a throughput of 85000 spheroids per microfluidic circuit per hour. The capability of automated spheroid generation is directly applicable to precision medicine and drug screening. Multiple cell lines were successfully tested, including cancer cell lines, co-cultures, and primary cells. The 3D-microtissues/spheroids were automatically assembled-incubated-retrieved with high viability for further drug screening analysis - the platform interfaces with standard labware.
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| 6. |
- Parker, Helen E., et al.
(författare)
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A Lab-in-a-Fiber optofluidic device using droplet microfluidics and laser-induced fluorescence for virus detection
- 2022
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Ingår i: Scientific Reports. - : Nature Research. - 2045-2322. ; 12:1
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Tidskriftsartikel (refereegranskat)abstract
- Microfluidics has emerged rapidly over the past 20 years and has been investigated for a variety of applications from life sciences to environmental monitoring. Although continuous-flow microfluidics is ubiquitous, segmented-flow or droplet microfluidics offers several attractive features. Droplets can be independently manipulated and analyzed with very high throughput. Typically, microfluidics is carried out within planar networks of microchannels, namely, microfluidic chips. We propose that fibers offer an interesting alternative format with key advantages for enhanced optical coupling. Herein, we demonstrate the generation of monodisperse droplets within a uniaxial optofluidic Lab-in-a-Fiber scheme. We combine droplet microfluidics with laser-induced fluorescence (LIF) detection achieved through the development of an optical side-coupling fiber, which we term a periscope fiber. This arrangement provides stable and compact alignment. Laser-induced fluorescence offers high sensitivity and low detection limits with a rapid response time making it an attractive detection method for in situ real-time measurements. We use the well-established fluorophore, fluorescein, to characterize the Lab-in-a-Fiber device and determine the generation of ∼ 0.9 nL droplets. We present characterization data of a range of fluorescein concentrations, establishing a limit of detection (LOD) of 10 nM fluorescein. Finally, we show that the device operates within a realistic and relevant fluorescence regime by detecting reverse-transcription loop-mediated isothermal amplification (RT-LAMP) products in the context of COVID-19 diagnostics. The device represents a step towards the development of a point-of-care droplet digital RT-LAMP platform. © 2022, The Author(s).
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| 7. |
- Parker, Helen E., et al.
(författare)
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Digital detection and quantification of SARS-CoV-2 in a droplet microfluidic all-fiber device
- 2021
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Ingår i: Proceedings MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. - : Chemical and Biological Microsystems Society. ; , s. 1047-1048
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Konferensbidrag (refereegranskat)abstract
- Silica fibers and capillaries offer opportunities for compact integration of optics with microfluidics while adding advantages such as; flexibility within a high aspect ratio format, uniaxial arrangements, and measurement-at-a-distance. Here, we describe droplet microfluidics-based nucleic acid detection of SARS-CoV-2 in a lab-in-a-fiber platform. The fiber component integrates three modules with key functions: droplet generation, incubation, and fluorescence detection. Within the scope of this work, we developed the component specifically to target the quantification of SARS-CoV-2 viral RNA through reverse-transcription loop-mediated isothermal amplification (RT-LAMP). The all-fiber component could successfully generate uniform droplets and differentiate pre-amplified positive LAMP reaction from negative sample.
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| 8. |
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| 9. |
- Parker, Helen E., et al.
(författare)
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Viral detection and quantification in a digital droplet microfluidic lab-in-a-fiber device
- 2021
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Ingår i: Proceedings of SPIE - The International Society for Optical Engineering. - : SPIE. - 9781510643802
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Konferensbidrag (refereegranskat)abstract
- In this work, we present the design and fabrication of a fiber device that performs digital droplet microfluidics for molecular diagnostics. A variety of fibers and capillaries were used to build three connected modules dedicated to droplet generation, incubation, and fluorescence detection which enables a uniaxial arrangement. This is in contrast to the traditional 2-dimensional lab-on-a-chip architecture. We characterize our fiber device using a fluorescein dilution series. Our observed detection limit is on the order of 10 nM fluorescein. We demonstrate our all-fiber device for the fluorescence readout after loop-mediated isothermal amplification (LAMP) of synthetic SARS-CoV-2. Our results suggest that this fiber device can successfully distinguish between positive and negative samples in molecular diagnostics. We propose that our fiber device offers benefits over microfluidic chip techniques such as easier optical integration, much simpler sample loading, and faster diagnosis with high specificity and sensitivity. Keywords: All-fiber device, microfluidics, optofluidics, loop-mediated isothermal amplification (LAMP), molecular diagnostics, SARS-CoV2.
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| 10. |
- Periyannan Rajeswari, Prem Kumar, et al.
(författare)
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Multiple pathogen biomarker detection using an encoded bead array in droplet PCR
- 2017
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Ingår i: Journal of Microbiological Methods. - : Elsevier. - 0167-7012 .- 1872-8359. ; 139, s. 22-28
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Tidskriftsartikel (refereegranskat)abstract
- We present a droplet PCR workflow for detection of multiple pathogen DNA biomarkers using fluorescent color coded Luminex beads. This strategy enables encoding of multiple singleplex droplet PCRs using a commercially available bead set of several hundred distinguishable fluorescence codes. This workflow provides scalability beyond the limited number offered by fluorescent detection probes such as TaqMan probes, commonly used in current multiplex droplet PCRs. The workflow was validated for three different Luminex bead sets coupled to target specific capture oligos to detect hybridization of three microorganisms infecting poultry: avian influenza, infectious laryngotracheitis virus and Campylobacter jejuni. In this assay, the target DNA was amplified with fluorescently labeled primers by PCR in parallel in monodisperse picoliter droplets, to avoid amplification bias. The color codes of the Luminex detection beads allowed concurrent and accurate classification of the different bead sets used in this assay. The hybridization assay detected target DNA of all three microorganisms with high specificity, from samples with average target concentration of a single DNA template molecule per droplet. This workflow demonstrates the possibility of increasing the droplet PCR assay detection panel to detect large numbers of targets in parallel, utilizing the scalability offered by the color-coded Luminex detection beads.
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