| 1. |
- Broman, Axel, et al.
(författare)
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Multinodal Acoustic Trapping Enables High Capacity and High Throughput Enrichment of Extracellular Vesicles and Microparticles in miRNA and MS Proteomics Studies
- 2021
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 1520-6882 .- 0003-2700. ; 93:8, s. 3929-3937
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Tidskriftsartikel (refereegranskat)abstract
- We report a new design of an acoustophoretic trapping device with significantly increased capacity and throughput, compared to current commercial acoustic trapping systems. Acoustic trapping enables nanoparticle and extracellular vesicle (EV) enrichment without ultracentrifugation. Current commercial acoustic trapping technology uses an acoustic single-node resonance and typically operates at flow rates <50 μL/min, which limits the processing of the larger samples. Here, we use a larger capillary that supports an acoustic multinode resonance, which increased the seed particle capacity 40 times and throughput 25-40 times compared to single-node systems. The resulting increase in capacity and throughput was demonstrated by isolation of nanogram amounts of microRNA from acoustically trapped urinary EVs within 10 min. Additionally, the improved trapping performance enabled isolation of extracellular vesicles for downstream mass spectrometry analysis. This was demonstrated by the differential protein abundance profiling of urine samples (1-3 mL), derived from the non-trapped versus trapped urine samples.
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| 2. |
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| 3. |
- Urbansky, Anke, et al.
(författare)
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Affinity-bead-mediated enrichment of CD8+ lymphocytes from peripheral blood progenitor cell products using acoustophoresis
- 2016
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Ingår i: Micromachines. - : MDPI AG. - 2072-666X. ; 7:6
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Tidskriftsartikel (refereegranskat)abstract
- Acoustophoresis is a technique that applies ultrasonic standing wave forces in a microchannel to sort cells depending on their physical properties in relation to the surrounding media. Cell handling and separation for research and clinical applications aims to efficiently separate specific cell populations. Here, we investigated the sorting of CD8 lymphocytes from peripheral blood progenitor cell (PBPC) products by affinity-bead-mediated acoustophoresis. PBPC samples were obtained from healthy donors (n = 4) and patients (n = 18). Mononuclear cells were labeled with anti-CD8-coated magnetic beads and sorted on an acoustophoretic microfluidic device and by standard magnetic cell sorting as a reference method. CD8 lymphocytes were acoustically sorted with a mean purity of 91% ± 8% and a median separation efficiency of 63% (range 15.1%-90.5%) as compared to magnetic sorting (purity 91% ± 14%, recovery 29% (range 5.1%-47.3%)). The viability as well as the proliferation capacity of sorted lymphocytes in the target fraction were unimpaired and, furthermore, hematopoietic progenitor cell assay revealed a preserved clonogenic capacity post-sorting. Bead-mediated acoustophoresis can, therefore, be utilized to efficiently sort less frequent CD8+ lymphocytes from PBPC products in a continuous flow mode while maintaining cell viability and functional capacity of both target and non-target fractions.
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| 4. |
- Olm, Franziska, et al.
(författare)
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Acoustophoresis enables the label-free separation of functionally different subsets of cultured bone marrow stromal cells
- 2021
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Ingår i: Cytometry. Part A : the journal of the International Society for Analytical Cytology. - : Wiley. - 1552-4930. ; 99:5, s. 476-487
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Tidskriftsartikel (refereegranskat)abstract
- Culture-expanded mesenchymal stromal cells (MSCs) are promising candidates for clinical cell-based therapies. MSC products are heterogeneous and we therefore investigated whether acoustophoresis, an ultrasound-based separation technology, could be used for the label-free enrichment of functionally different MSC populations. Acoustophoresis uses an ultrasonic standing wave in a microchannel which differentially affects the movement of cells depending on their acoustophysical properties, such as size, density, and compressibility. Human bone marrow MSCs were generated by standard adherent culture in xenofree medium and separated by microchip acoustophoresis. MSCs with up to 20% higher proliferation and 1.7-fold increased clonogenic potential were enriched in the side outlet of the chip compared to the input sample. These cells were significantly smaller (average diameter 14.5 ± 0.4 μm) compared to the center outlet fraction (average diameter 17.1 ± 0.6 μm) and expressed higher levels of genes related to proliferation and stem cell properties (i.e. Ki-67 (1.9-fold), Nanog1 (6.65-fold), Oct4 (2.9-fold), and CXCL12 (1.8-fold), n = 3) in the side outlet compared to input. Fractions of MSCs in G0 /G1 cell cycle phase were significantly enriched in the side fraction and an up to 2.8-fold increase of cells in S/G2 /M phases were observed in center fractions compared to side fractions and 1.3-fold increased compared to the input sample. Acoustophoresis did not compromise MSC phenotype, proliferation, clonogenic capacity, and viability (generally 87-98%), nor did it affect differentiation or immunomodulatory capacities. These results demonstrate that label-free acoustic separation can enrich functionally different MSC subsets which can potentially be employed to produce better-defined stromal cell products from cultured MSCs. Hence, acoustophoresis is a potentially promising separation technology to provide improved cell products for research and possible future clinical use.
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| 5. |
- Urbansky, Anke, et al.
(författare)
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Label-free separation of leukocyte subpopulations using high throughput multiplex acoustophoresis
- 2019
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 19:8, s. 1406-1416
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Tidskriftsartikel (refereegranskat)abstract
- Multiplex separation of mixed cell samples is required in a variety of clinical and research applications. Herein, we present an acoustic microchip with multiple outlets and integrated pre-alignment channel to enable high performance and label-free separation of three different cell or particle fractions simultaneously at high sample throughput. By implementing a new cooling system for rigorous temperature control and minimal acoustic energy losses, we were able to operate the system isothermally and sort suspensions of 3, 5 and 7 μm beads with high efficiencies (>95.4%) and purities (>96.3%) at flow rates up to 500 μL min -1 corresponding to a throughput of ∼2.5 × 10 6 beads per min. Also, human viable white blood cells were successfully fractionated into lymphocytes, monocytes and granulocytes with high purities of 96.5 ± 1.6%, 71.8 ± 10.1% and 98.8 ± 0.5%, respectively, as well as high efficiencies (96.8 ± 3.3%, 66.7 ± 3.2% and 99.0 ± 0.7%) at flow rates up to 100 μL min -1 (∼100000 cells per min). By increasing the flow rate up to 300 μL min -1 (∼300000 cells per min) both lymphocytes and granulocytes were still recovered with high purities (92.8 ± 1.9%, 98.2 ± 1.0%), whereas the monocyte purity decreased to 20.9 ± 10.3%. The proposed isothermal multiplex acoustophoresis platform offers efficient fractionation of complex samples in a label-free and continuous manner at thus far unreached high sample throughput rates.
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| 6. |
- Barnkob, Rune, et al.
(författare)
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Measuring density and compressibility of white blood cells and prostate cancer cells by microchannel acoustophoresis
- 2011
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Ingår i: 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2011, MicroTAS 2011. - 9781618395955 ; 1, s. 127-129
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Konferensbidrag (refereegranskat)abstract
- We present a novel method for the determination of density and compressibility of individual particles and cells undergoing microchannel acoustophoresis in an arbitrary 2D acoustic field. Our method is a critical advancement within acoustophoretic separation of biological cells, as the ability to determine the density and compressibility of individual cells enables the prediction and alteration of the separation outcome for a given cell mixture. We apply the method on white blood cells (WBCs) and DU145 prostate cancer cells (DUCs) aiming to improve isolation of circulating tumor cells from blood, an emerging tool in the monitoring and characterizing of metastatic cancer.
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| 7. |
- Järås, Kerstin, et al.
(författare)
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PSA quantification of 80 plasma samples from the clinical routine using antibody microarrays
- 2009
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Ingår i: Proceedings of Conference, MicroTAS 2009 - The 13th International Conference on Miniaturized Systems for Chemistry and Life Sciences. - 9780979806421 ; , s. 457-459
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Konferensbidrag (refereegranskat)abstract
- Antibody microarrays are becoming increasingly established in clinical studies. However, the arrays are seldom used in a quantitative approach but rather for detecting up or down regulated proteins. In this study we describe a microarray procedure being standardized by placing the in-house developed porous silicon surfaces into a commercially available 96 well microtiter plate for analyzing the PSA concentrationsof 80 EDTA-plasma samples obtained from patients undergoing clinical PSAtesting.
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| 8. |
- Lenshof, Andreas, et al.
(författare)
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Acoustic whole blood plasmapheresis chip for PSA microarray diagnostics
- 2009
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Ingår i: Proceedings of Conference, MicroTAS 2009 : The 13th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The 13th International Conference on Miniaturized Systems for Chemistry and Life Sciences. - 9780979806421 ; , s. 1330-1332
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Konferensbidrag (refereegranskat)abstract
- An acoustic device for generating high quality blood plasma for PSA microarray diagnostics is presented.
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| 9. |
- Lenshof, Andreas, et al.
(författare)
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High performance multiplex acoustophoresis for WBC subpopulation isolation
- 2017
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Ingår i: MicroTAS 2017 : Savannah, Georgia, USA - Savannah, Georgia, USA. - 1556-5904. - 9780692941836 ; 2017, s. 1385-1386
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Konferensbidrag (refereegranskat)abstract
- Recently, acoustophoresis has been used to fractionate white blood cells (WBC) into subpopulations, Grenvall et al. [1]. However, at a sample throughput of 8-10 μl/min the separation has limited bioanalytical application. In order to substantially increase throughput, we have redesigned and developed a new separation system that enables unmatched WBC separation performance at a volume throughput of 200μl/min and a cell concentration of 106 cells/ml.
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| 10. |
- Rezeli, Melinda, et al.
(författare)
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Comparative Proteomic Analysis of Extracellular Vesicles Isolated by Acoustic Trapping or Differential Centrifugation
- 2016
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 88:17, s. 8577-8586
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Tidskriftsartikel (refereegranskat)abstract
- Extracellular vesicles (ECVs), including microparticles and exosomes, are submicrometer membrane vesicles released by diverse cell types upon activation or stress. Circulating ECVs are potential reservoirs of disease biomarkers, and the complexity of these vesicles is significantly lower compared to their source, blood plasma, which makes ECV-based biomarker studies more promising. Proteomic profiling of ECVs is important not only to discover new diagnostic or prognostic markers but also to understand their roles in biological function. In the current study, we investigated the protein composition of plasma-derived ECVs isolated by acoustic seed trapping. Additionally, the protein composition of ECVs isolated with acoustic trapping was compared to that isolated with a conventional differential centrifugation protocol. Finally, the proteome of ECVs originating from ST-elevation myocardial infarction patients was compared with that of healthy controls using label-free LC-MS quantification. The acoustic trapping platform allows rapid and automated preparation of ECVs from small sample volumes, which are therefore well-suited for biobank repositories. We found that the protein composition of trapped ECVs is very similar to that isolated by the conventional differential centrifugation method.
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