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On-chip ultrasonic sample preparation for cell based assays
KTH, School of Engineering Sciences (SCI), Applied Physics.ORCID iD: 0000-0003-0064-0086
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Engineering Sciences (SCI), Applied Physics.
2015 (English)In: RSC Advances, ISSN 2046-2069, E-ISSN 2046-2069, Vol. 5, no 91, 74304-74311 p.Article in journal (Refereed) Published
Abstract [en]

We demonstrate an acoustophoresis method for size-based separation, isolation, up-concentration and trapping of cells that can be used for on-chip sample preparation combined with high resolution imaging for cell-based assays. The method combines three frequency-specific acoustophoresis functions in a sequence by actuating three separate channel zones simultaneously: zones for pre-alignment, size-based separation, and trapping. We characterize the mutual interference between the acoustic radiation forces between the different zones by measuring the spatial distribution of the acoustic energy density during different schemes of ultrasonic actuation, and use this information for optimizing the driving frequencies and voltages of the three utilized ultrasonic transducers attached to the chip, and the flow rates of the pumps. By the use of hydrodynamic defocusing of the pre-aligned cells in the separation zone, a cell population from a complex sample can be isolated and trapped with very high purity, followed by dynamic fluorescence analysis. We exemplify the method's potential by isolating A549 lung cancer cells from red blood cells with 100% purity, 92% separation efficiency, and 93% trapping efficiency resulting in a 130× up-concentration factor during 15 minutes of continuous sample processing through the chip. Furthermore, we demonstrate an on-chip fluorescence assay of the isolated cancer cells by monitoring the dynamic uptake and release of a fluorescence probe in individual trapped cells. The ability to combine isolation of individual cells from a complex sample with high-resolution image analysis holds great promise for applications in cellular and molecular diagnostics.

Place, publisher, year, edition, pages
RSC Publishing, 2015. Vol. 5, no 91, 74304-74311 p.
National Category
Chemical Sciences
URN: urn:nbn:se:kth:diva-166823DOI: 10.1039/c5ra16865aISI: 000361116500020ScopusID: 2-s2.0-84941242035OAI: diva2:812495

Updated from manuscript to article.

QC 20151008

Available from: 2015-05-19 Created: 2015-05-19 Last updated: 2015-11-05Bibliographically approved
In thesis
1. On-chip Ultrasonic Sample Preparation
Open this publication in new window or tab >>On-chip Ultrasonic Sample Preparation
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Acoustofluidics has become a well-established technology in the lab-on-a-chip scientific community. The technology involves primarily the manipulation of fluids and/or particles in microfluidic systems. It is used today for variety of applications such as handling, sorting, washing and separation of cells or micro-particles, and for mixing and pumping of fluids. When such manipulation functions are integrated in micro-devices, the technology has been used for clinical sample preparation as well as for studying various fundamental bio-related questions.

In this doctoral thesis, we have developed different acoustic methods and micro-devices with the aim to create a multi-functional sample preparation platform. We introduced a simple method for in-situ measurements of acoustic energy densities inside a microfluidic channel, from which acoustic pressure amplitudes can be extracted. The method has been used for determining the magnitude of acoustic radiation forces acting on suspended particles and cells inside an acoustofluidic system. For optimization of acoustophoresis (i.e. manipulation of particles into the nodes of standing waves), we have investigated different designs of ultrasonic transducers based on tunable-angle wedges and backing layers attached to glass-silicon microfluidic chips. Furthermore, we have investigated the implementation of frequency-modulated actuation methodology combined with broadbanded ultrasonic transducers, and the implementation of multiple ultrasonic manipulation functions localized to spatially separated zones in a complex microchannel network. We demonstrate two different bio-applications useful for multi-step and multi-functional sample preparation. First, we demonstrate a micro-device for size-based separation, isolation and up-concentration of cells, followed by microscopy-based dynamic monitoring of individual cell properties when introducing different reagents. This holds great promise for use in cellular and molecular diagnostics. Second, we demonstrate an acoustic method for micro-vortexing in µL-volume reaction chambers in disposable polymer chips. The method is used for fast mixing of fluids, for disaggregating and re-suspending magnetically trapped and clumped micro-beads, and for cell lysis followed by DNA extraction. Finally, we demonstrate a temperature-controlled device compatible with high-acoustic-pressure (1 MPa) ultrasonic manipulation of cells, and we demonstrate that cells can be exposed to standing-wave ultrasound at 1 MPa for one hour without compromising the cell viability.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2015. ix, 65 p.
TRITA-FYS, ISSN 0280-316X ; 15:21
Ultrasound, Sample preparation, Particle manipulation
National Category
Other Physics Topics
Research subject
Physics; Biological Physics
urn:nbn:se:kth:diva-166746 (URN)978-91-7595-529-2 (ISBN)
Public defence
2015-06-05, FD5 AlbaNova University centrum,, Roslagstullsbacken 21, KTH, Stockholm, 07:14 (English)

QC 20150519

Available from: 2015-05-19 Created: 2015-05-15 Last updated: 2015-06-04Bibliographically approved

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