| 1. |
- Urbansky, Anke, et al.
(author)
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Affinity-bead-mediated enrichment of CD8+ lymphocytes from peripheral blood progenitor cell products using acoustophoresis
- 2016
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In: Micromachines. - : MDPI AG. - 2072-666X. ; 7:6
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Journal article (peer-reviewed)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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| 2. |
- Urbansky, Anke, et al.
(author)
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Label-free separation of leukocyte subpopulations using high throughput multiplex acoustophoresis
- 2019
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In: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 19:8, s. 1406-1416
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Journal article (peer-reviewed)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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| 3. |
- Lenshof, Andreas, et al.
(author)
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High performance multiplex acoustophoresis for WBC subpopulation isolation
- 2017
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In: MicroTAS 2017 : Savannah, Georgia, USA - Savannah, Georgia, USA. - 1556-5904. - 9780692941836 ; 2017, s. 1385-1386
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Conference paper (peer-reviewed)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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| 4. |
- Urbansky, Anke, et al.
(author)
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Label-free acoustophoretic enrichment of mononuclear cells from blood
- 2017
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In: MicroTAS 2017 : Savannah, Georgia, USA - Savannah, Georgia, USA. - 1556-5904. - 9780692941836 ; 2017, s. 1309-1310
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Conference paper (peer-reviewed)abstract
- We are reporting an efficient, label-free and continuous separation of mononuclear cells (MNC), from blood using microchip acoustophoresis. In standard PBS buffer, MNCs and red blood cells (RBC) display overlapping acoustophoretic mobilities which compromise separation of these cell types from each other. In this paper we capitalize on the fact that MNC and RBC display different acoustophysical properties. By optimizing the buffer conditions and thereby changing the acoustic contrast factor, and hence the acoustophoretic mobility of the cells, a 2800-fold enrichment of MNCs vs. RBCs with MNC recoveries up to 88% was accomplished.
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