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Iranmanesh, I. S. (2015). On-chip Ultrasonic Sample Preparation. (Doctoral dissertation). Stockholm: KTH Royal Institute of Technology
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. p. ix, 65
Series
TRITA-FYS, ISSN 0280-316X ; 15:21
Keywords
Ultrasound, Sample preparation, Particle manipulation
National Category
Other Physics Topics
Research subject
Physics; Biological Physics
Identifiers
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)
Opponent
Supervisors
Note

QC 20150519

Available from: 2015-05-19 Created: 2015-05-15 Last updated: 2015-06-04Bibliographically approved
Iranmanesh, I. S., Ramachandraiah, H., Russom, A. & Wiklund, M. (2015). On-chip ultrasonic sample preparation for cell based assays. RSC Advances, 5(91), 74304-74311
Open this publication in new window or tab >>On-chip ultrasonic sample preparation for cell based assays
2015 (English)In: RSC Advances, ISSN 2046-2069, E-ISSN 2046-2069, Vol. 5, no 91, p. 74304-74311Article 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
National Category
Chemical Sciences
Identifiers
urn:nbn:se:kth:diva-166823 (URN)10.1039/c5ra16865a (DOI)000361116500020 ()2-s2.0-84941242035 (Scopus ID)
Note

Updated from manuscript to article.

QC 20151008

Available from: 2015-05-19 Created: 2015-05-19 Last updated: 2017-12-04Bibliographically approved
Ohlin, M., Iranmanesh, I. S., Christakou, A. E. & Wiklund, M. (2015). Temperature-controlled MPa-pressure ultrasonic cell manipulation in a microfluidic chip. Lab on a Chip, 15(16), 3341-3349
Open this publication in new window or tab >>Temperature-controlled MPa-pressure ultrasonic cell manipulation in a microfluidic chip
2015 (English)In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 15, no 16, p. 3341-3349Article in journal (Refereed) Published
Abstract [en]

We study the temperature-independent impact on cell viability of relevant physical parameters during long-term, high-acoustic-pressure ultrasonic exposure in a microfluidic chip designed for ultrasonic-standing-wave trapping and aggregation of cells. We use a light-intensity method and 5 mum polymer beads for accurate acoustic pressure calibration before injecting cells into the device, and we monitor the viability of A549 lung cancer cells trapped during one hour in an ultrasonic standing wave with 1 MPa pressure amplitude. The microfluidic chip is actuated by a novel temperature-controlled ultrasonic transducer capable of keeping the temperature stable around 37 °C with an accuracy better than ±0.2 °C, independently on the ultrasonic power and heat produced by the system, thereby decoupling any temperature effect from other relevant effects on cells caused by the high-pressure acoustic field. We demonstrate that frequency-modulated ultrasonic actuation can produce acoustic pressures of equally high magnitudes as with single-frequency actuation, and we show that A549 lung cancer cells can be exposed to 1 MPa standing-wave acoustic pressure amplitudes for one hour without compromising cell viability. At this pressure level, we also measure the acoustic streaming induced around the trapped cell aggregate, and conclude that cell viability is not affected by streaming velocities of the order of 100 mum s(-1). Our results are important when implementing acoustophoresis methods in various clinical and biomedical applications.

National Category
Engineering and Technology
Identifiers
urn:nbn:se:kth:diva-166828 (URN)10.1039/c5lc00490j (DOI)000358609500011 ()26156858 (PubMedID)2-s2.0-84938342105 (Scopus ID)
Note

Updated from "Manuscript" to "Article". QC 20150814

Available from: 2015-05-19 Created: 2015-05-19 Last updated: 2020-01-29Bibliographically approved
Wiklund, M., Christakou, A. E., Ohlin, M., Iranmanesh, I., Frisk, T., Vanherberghen, B. & Önfelt, B. (2014). Ultrasound-Induced Cell-Cell Interaction Studies in a Multi-Well Microplate. Micromachines, 5(1), 27-49
Open this publication in new window or tab >>Ultrasound-Induced Cell-Cell Interaction Studies in a Multi-Well Microplate
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2014 (English)In: Micromachines, ISSN 2072-666X, E-ISSN 2072-666X, Vol. 5, no 1, p. 27-49Article, review/survey (Refereed) Published
Abstract [en]

This review describes the use of ultrasound for inducing and retaining cell-cell contact in multi-well microplates combined with live-cell fluorescence microscopy. This platform has been used for studying the interaction between natural killer (NK) cells and cancer cells at the level of individual cells. The review includes basic principles of ultrasonic particle manipulation, design criteria when building a multi-well microplate device for this purpose, biocompatibility aspects, and finally, two examples of biological applications: Dynamic imaging of the inhibitory immune synapse, and studies of the heterogeneity in killing dynamics of NK cells interacting with cancer cells.

Keywords
ultrasound, lab-on-a-chip, acoustofluidics, acoustic trapping, natural killer cells
National Category
Nano Technology Biological Sciences
Identifiers
urn:nbn:se:kth:diva-144959 (URN)10.3390/mi5010027 (DOI)000333674700003 ()2-s2.0-84902585350 (Scopus ID)
Funder
Swedish Foundation for Strategic Research EU, FP7, Seventh Framework ProgrammeSwedish Research Council
Note

QC 20140505

Available from: 2014-05-05 Created: 2014-05-05 Last updated: 2020-01-29Bibliographically approved
Iranmanesh, I., Barnkob, R., Bruus, H. & Wiklund, M. (2013). Magnitude and variance of acoustic energy density in microchannel acoustophoresis: Comparison between single-frequency and frequency-modulated actuation. In: 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013: . Paper presented at 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013; Freiburg; Germany (pp. 1400-1402). , 3
Open this publication in new window or tab >>Magnitude and variance of acoustic energy density in microchannel acoustophoresis: Comparison between single-frequency and frequency-modulated actuation
2013 (English)In: 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013, 2013, Vol. 3, p. 1400-1402Conference paper, Published paper (Refereed)
Abstract [en]

Using a novel light-intensity method, we quantify for the first time the magnitude and spatial variance in acoustic energy density along a microchannel during acoustophoretic focusing of particles with frequency-modulated ultrasound. We compare the distribution in energy density between single-frequency (SF) and frequency-modulation (FM) actuation along the microchannel. In addition, we analyze the field uniformity for the two actuation approaches (SF and FM) by measuring the deviation of the final particle pattern from an ideal straight line. We conclude that the magnitude of the energy density for FM actuation is of the same order of magnitude as for SF actuation, but with much less spatial variance.

Keywords
Acoustic energy density, Acoustophoresis, Frequency modulation, Microchannel
National Category
Fluid Mechanics and Acoustics
Identifiers
urn:nbn:se:kth:diva-168793 (URN)2-s2.0-84907311740 (Scopus ID)978-163266624-6 (ISBN)
Conference
17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013; Freiburg; Germany
Note

QC 20150611

Available from: 2015-06-11 Created: 2015-06-09 Last updated: 2015-06-11Bibliographically approved
Wiklund, M., Christakou, A. E., Iranmanesh, I., Ohlin, M., Russom, A. & Önfelt, B. (2013). On-chip acoustic sample preparation for cell studies and diagnostics. In: Proceedings of Meetings on Acoustics: Volume 19, 2013. Paper presented at 21st International Congress on Acoustics, ICA 2013 - 165th Meeting of the Acoustical Society of America; Montreal, QC; Canada; 2 June 2013 through 7 June 2013 (pp. 1-3). Acoustical Society of America (ASA)
Open this publication in new window or tab >>On-chip acoustic sample preparation for cell studies and diagnostics
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2013 (English)In: Proceedings of Meetings on Acoustics: Volume 19, 2013, Acoustical Society of America (ASA), 2013, p. 1-3Conference paper, Published paper (Refereed)
Abstract [en]

We describe a novel platform for acoustic sample preparation in microchannels and microplates. The utilized method is based on generating a multitude of acoustic resonances at a set of different frequencies in microstructures, in order to accurately control the migration and positioning of particles and cells suspended in fluid channels and chambers. The actuation frequencies range from 30 kHz to 7 MHz, which are applied simultaneously and/or in sweeps. We present two devices: A closed microfluidic chip designed for pre-alignment, size-based separation, isolation, up-concentration and lysis of cells, and an open multi-well microplate designed for parallel aggregation and positioning of cells. Both devices in the platform are compatible with high-resolution live-cell microscopy, which is used for fluorescence-based optical characterization. Two bioapplications are demonstrated for each of the devices: The first device is used for size-selective cell isolation and lysis for DNA-based diagnostics, and the second device is used for quantifying the heterogeneity in cytotoxic response of natural killer cells interacting with cancer cells.

Place, publisher, year, edition, pages
Acoustical Society of America (ASA), 2013
Series
Proceedings of Meetings on Acoustics, ISSN 1939-800X ; 19
Keywords
Acoustic resonance, Bioapplications, Cytotoxic response, Different frequency, Microfluidic chip, Natural killer cells, Optical characterization, Sample preparation
National Category
Biological Sciences
Identifiers
urn:nbn:se:kth:diva-134244 (URN)10.1121/1.4800637 (DOI)2-s2.0-84878982614 (Scopus ID)
Conference
21st International Congress on Acoustics, ICA 2013 - 165th Meeting of the Acoustical Society of America; Montreal, QC; Canada; 2 June 2013 through 7 June 2013
Note

QC 20131121

Available from: 2013-11-21 Created: 2013-11-20 Last updated: 2020-01-29Bibliographically approved
Iranmanesh, I. S., Barnkob, R., Bruus, H. & Wiklund, M. (2013). Tunable-angle wedge transducer for improved acoustophoretic control in a microfluidic chip. Journal of Micromechanics and Microengineering, 23(10), 105002
Open this publication in new window or tab >>Tunable-angle wedge transducer for improved acoustophoretic control in a microfluidic chip
2013 (English)In: Journal of Micromechanics and Microengineering, ISSN 0960-1317, E-ISSN 1361-6439, Vol. 23, no 10, p. 105002-Article in journal (Refereed) Published
Abstract [en]

We present a tunable-angle wedge ultrasound transducer for improved control of microparticle acoustophoresis in a microfluidic chip. The transducer is investigated by analyzing the pattern of aligned particles and induced acoustic energy density while varying the transducer geometry, transducer coupling angle, and transducer actuation method (single-frequency actuation or frequency-modulation actuation). The energy-density analysis is based on measuring the transmitted light intensity through a microfluidic channel filled with a suspension of 5 mu m diameter beads and the results with the tunable-angle transducer are compared with the results from actuation by a standard planar transducer in order to decouple the influence from change in coupling angle and change in transducer geometry. We find in this work that the transducer coupling angle is the more important parameter compared to the concomitant change in geometry and that the coupling angle may be used as an additional tuning parameter for improved acoustophoretic control with single-frequency actuation. Further, we find that frequency-modulation actuation is suitable for diminishing such tuning effects and that it is a robust method to produce uniform particle patterns with average acoustic energy densities comparable to those obtained using single-frequency actuation.

Keywords
Acoustic Radiation Force, Small Particles, Cell, Manipulation, Resonators, Channels, Field
National Category
Engineering and Technology
Identifiers
urn:nbn:se:kth:diva-131713 (URN)10.1088/0960-1317/23/10/105002 (DOI)000324672700003 ()2-s2.0-84884878282 (Scopus ID)
Funder
Swedish Research Council, 2011-5230EU, FP7, Seventh Framework Programme
Note

QC 20131018

Available from: 2013-10-18 Created: 2013-10-17 Last updated: 2017-12-06Bibliographically approved
Barnkob, R., Iranmanesh, I., Wiklund, M. & Bruus, H. (2012). Measuring acoustic energy density in microchannel acoustophoresis using a simple and rapid light-intensity method. Lab on a Chip, 12(13), 2337-2344
Open this publication in new window or tab >>Measuring acoustic energy density in microchannel acoustophoresis using a simple and rapid light-intensity method
2012 (English)In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 12, no 13, p. 2337-2344Article in journal (Refereed) Published
Abstract [en]

We present a simple and rapid method for measuring the acoustic energy density in microchannel acoustophoresis based on light-intensity measurements of a suspension of particles. The method relies on the assumption that each particle in the suspension undergoes single-particle acoustophoresis. It is validated by the single-particle tracking method, and we show by proper re-scaling that the re-scaled light intensity plotted versus re-scaled time falls on a universal curve. The method allows for analysis of moderate-resolution images in the concentration range encountered in typical experiments, and it is an attractive alternative to particle tracking and particle image velocimetry for quantifying acoustophoretic performance in microchannels.

Keywords
Ultrasonic Standing-Wave, Chip, Cell, Manipulation, Suspensions, Trap
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:kth:diva-98959 (URN)10.1039/c2lc40120g (DOI)000305288600009 ()2-s2.0-84862209428 (Scopus ID)
Funder
Swedish Research Council, 2011-5230
Note

QC 20120710

Available from: 2012-07-10 Created: 2012-07-05 Last updated: 2017-12-07Bibliographically approved
Svennebring, J., Manneberg, O., Iranmanesh, I., Hertz, H. M. & Wiklund, M. (2009). Ultrasonic manipulation in microfluidic systems: Selective cell handling and characterization. Paper presented at 7th USWNet Meeting: Unidirectional motion produced by vibrating fields for cell/particle and fluid control. Stockholm, Sweden. Nov. 30th - Dec. 1st, 2009.
Open this publication in new window or tab >>Ultrasonic manipulation in microfluidic systems: Selective cell handling and characterization
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2009 (English)Conference paper, Published paper (Other academic)
National Category
Other Physics Topics
Identifiers
urn:nbn:se:kth:diva-90429 (URN)
Conference
7th USWNet Meeting: Unidirectional motion produced by vibrating fields for cell/particle and fluid control. Stockholm, Sweden. Nov. 30th - Dec. 1st, 2009
Note
QC 20120417Available from: 2012-02-23 Created: 2012-02-23 Last updated: 2012-04-17Bibliographically approved
Iranmanesh, I. S., Ohlin, M., Ramachandraiah, H., Russom, A. & Wiklund, M. Acoustic micro-vortexing of fluids, beads and cells in disposable microfluidic chips.
Open this publication in new window or tab >>Acoustic micro-vortexing of fluids, beads and cells in disposable microfluidic chips
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(English)Manuscript (preprint) (Other academic)
National Category
Engineering and Technology
Identifiers
urn:nbn:se:kth:diva-166825 (URN)
Note

QS 2015

Available from: 2015-05-19 Created: 2015-05-19 Last updated: 2020-01-29Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0003-0064-0086

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