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MicroBubble Activated Acoustic Cell Sorting: BAACS
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi. KTH, Centra, Science for Life Laboratory, SciLifeLab. mafaridi@kth.se. (Clinical Microfluidics)ORCID-id: 0000-0003-1176-0905
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi. KTH, Centra, Science for Life Laboratory, SciLifeLab. (Clinical Microfluidics)ORCID-id: 0000-0001-5199-0663
KTH, Skolan för teknikvetenskap (SCI), Tillämpad fysik.ORCID-id: 0000-0003-0064-0086
KTH, Skolan för teknik och hälsa (STH), Medicinsk teknik, Medicinsk bildteknik.ORCID-id: 0000-0002-3699-396X
Visa övriga samt affilieringar
(Engelska)Ingår i: Biomedical microdevices (Print), ISSN 1387-2176, E-ISSN 1572-8781Artikel i tidskrift (Refereegranskat) Submitted
Abstract [en]

Acoustophoresis, the ability to acoustically manipulate particles and cells inside a microfluidic channel, is a critical enabling technology for cell-sorting applications. However, one of the major impediments for routine use of acoustophoresis at clinical laboratory has been the reliance on the inherent physical properties of cells for separation. Here, we present a microfluidic-based microBubble-Activated Acoustic Cell Sorting (BAACS) method that rely on the specific binding of target cells to microbubbles conjugated with specific antibodies on their surface for continuous cell separation using ultrasonic standing wave. In acoustophoresis, cells being positive acoustic contrast particles migrate to pressure nodes. On the contrary we show that air-filled polymer-shelled microbubbles being strong negative acoustic contrast particles migrate to pressure antinodes at acoustic pressure amplitudes as low as 60 kPa. As a proof of principle, using the BAACS strategy, we demonstrate the separation of cancer cell line in a suspension with better than 75% efficiency. Moreover, 100% of the microbubble-cell conjugates migrated to the anti-node. Hence a better upstream affinity-capture has the potential to provide higher sorting efficiency. The BAACS technique may potentially provide a simplistic approach for similar sized selective isolation of cells, and is suited for applications in point of care.

Nyckelord [en]
Cell sorting, acoustophoresis, microbubble, contrast agent, microfluidic separation
Nationell ämneskategori
Medicinsk bioteknologi
Identifikatorer
URN: urn:nbn:se:kth:diva-200302OAI: oai:DiVA.org:kth-200302DiVA, id: diva2:1068017
Forskningsfinansiär
EU, FP7, Sjunde ramprogrammet, 115153
Anmärkning

QC 20170124

Tillgänglig från: 2017-01-24 Skapad: 2017-01-24 Senast uppdaterad: 2017-11-29Bibliografiskt granskad
Ingår i avhandling
1. Bioparticle Manipulation using Acoustophoresis and Inertial Microfluidics
Öppna denna publikation i ny flik eller fönster >>Bioparticle Manipulation using Acoustophoresis and Inertial Microfluidics
2017 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Despite the many promising advances made in microfluidics, sample preparation remains the single largest challenge and bottleneck in the field of miniaturised diagnostics. This thesis is focused on the development of sample preparation methods using active and passive particle manipulation techniques for point of care diagnostic applications. The active technique is based on acoustophoresis (acoustic manipulation) while the passive method is based on inertial microfluidics (hydrodynamic manipulation). In paper I, acoustic capillary-based cavity resonator was used to study aggregation of silica and polystyrene particles. We found that silica particles show faster aggregation time (5.5 times) and larger average area of aggregates (3.4 times) in comparison to polystyrene particles under the same actuation procedure. The silica particles were then used for acoustic based bacteria up-concentration. In paper II, a microfluidic-based microbubbles activated acoustic cell sorting technique was developed for affinity based cell separation. As a proof of principle, separation of cancer cell line in a suspension with better than 75% efficiency is demonstrated. For the passive sample preparation, inertial and elasto-inertial microfluidic approach that uses geometry-induced hydrodynamic forces for continuous size-based sorting of particles in a flow-through fashion were studied and applied for blood processing (paper III-V). In paper III, a simple ushaped curved channel was used for inertial microfluidics based enrichment of white blood cells from diluted whole blood. A filtration efficiency of 78% was achieved at a flow rate of 2.2 ml/min. In paper IV, elasto-inertial microfluidics where viscoelastic flow enables size-based migration of cells into a non- Newtonian solution, was used to continuously separate bacteria from unprocessed whole blood for sepsis diagnostics. Bacteria were continuously separated at an efficiency of 76% from undiluted whole blood sample. Finally, in paper V, the inertial and elasto-inertial techniques were combined with a detection platform to demonstrate an integrated miniaturized flow cytometer. The all-optical-fiber technology based system allows for simultaneous measurements of fluorescent and scattering data at 2500 particles/s. The use of inertial and acoustic techniques for sample preparation and development of an integrated detection platform may allow for further development and realization of point of care testing (POCT) systems.

Ort, förlag, år, upplaga, sidor
Stockholm, Sweden: Kungliga Tekniska högskolan, 2017. s. 68
Serie
TRITA-BIO-Report, ISSN 1654-2312 ; 2017:4
Nationell ämneskategori
Medicinsk bioteknologi
Forskningsämne
Bioteknologi
Identifikatorer
urn:nbn:se:kth:diva-200304 (URN)978-91-7729-264-7 (ISBN)
Disputation
2017-02-16, Gard-Aulan, Nobels vägen 18, Solna, Stockholm, 10:00 (Engelska)
Opponent
Handledare
Anmärkning

QC 20170124

Tillgänglig från: 2017-01-24 Skapad: 2017-01-24 Senast uppdaterad: 2017-01-24Bibliografiskt granskad
2. Microfluidic based isolation of circulating tumor cells from whole blood for cancer diagnostics
Öppna denna publikation i ny flik eller fönster >>Microfluidic based isolation of circulating tumor cells from whole blood for cancer diagnostics
2017 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Detection of circulating tumor cells (CTC) in peripheral blood is indicative of early recognition of tumor progression and such an important biomarker for early diagnosis, staging, monitoring and prognosis of cancer. However, CTC are found in very low concentrations and reliable isolation of these rare cells is challenging. Microfluidics enables precise manipulation of fluids and cells and is ideal for cell sorting methods for clinical diagnostics. The thesis contributes towards the development of microfluidic based CTC isolation methods from peripheral blood. The methods are based on size and immunoaffinity. The first part of the thesis describes the phenomenon of inertial focusing for size based cell separation at high throughputs. In paper 1, we demonstrate continuous filtration of leukocytes from diluted blood, with an efficiency of 78% at a flow rate of 2.2ml/min. In the paper 2, separation of total and subpopulation of leukocytes with a purity of 86% for granulocytes and 91% for lymphocytes is demonstrated. Furthermore, cancer cells spiked into whole blood could be separated at a yield of 88%. Finally, in paper 3 and 4 we unravel parts of the unexplored elasto-inertial microfluidics and was utilized to precisely focus the cells, as part of an integrated optofluidic micro flow cytometer device, capable to simultaneously measure fluorescence and scattering of cells and particles at a rate of 2500 particles/sec (paper 4). Second part of the thesis focuses on acoustophoresis. In (paper 5), a multifunctional acoustic microdevice was developed for isolation of cancer cells from red blood cells with a separation efficiency of 92.4% and trapping efficiency of 93%. In (paper 6), microbubbles activated acoustic cell sorter was developed for affinity based cell separation. As a proof of principle, cancer cells in a suspension were separated at an efficiency of 75%. In the third part, using cellulose nano fibrils (paper 7), we demonstrate efficiently capture and release of cancer cells at a release efficiency of 95%. Finally, a novel, single step self-assembly of spider silk proteins is introduced inside microfluidic channels for effective capture of cancer cells with 85% capture efficiency and subsequent release of captured cells with 95% release efficiency (paper 8). The novel recombinant silk modified microfluidic device was validated using pancreatic cancer patients. In summary, we have developed different microfluidic based isolation technologies for the capture and characterization of CTC.

Ort, förlag, år, upplaga, sidor
stockholm: KTH Royal Institute of Technology, 2017. s. 109
Serie
TRITA-BIO-Report, ISSN 1654-2312 ; 2017:7
Nationell ämneskategori
Medicinteknik Medicinsk bioteknologi
Forskningsämne
Bioteknologi
Identifikatorer
urn:nbn:se:kth:diva-203889 (URN)978-91-7729-311-8 (ISBN)
Disputation
2017-04-13, Gardaulan, Folkhälsomyndigheten, Nobels väg 18, Solna, Stockholm, 10:00 (Engelska)
Opponent
Handledare
Forskningsfinansiär
EU, FP7, Sjunde ramprogrammetBarncancerfondenVetenskapsrådet
Anmärkning

QC 20170321

Tillgänglig från: 2017-03-20 Skapad: 2017-03-20 Senast uppdaterad: 2017-03-23Bibliografiskt granskad

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Grishenkov, DmitryWiklund, Martin

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Faridi, Muhammad AsimRamachandraiah, HarishaIranmanesh, Ida SadatGrishenkov, DmitryWiklund, MartinRussom, Aman
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Proteomik och nanobioteknologiScience for Life Laboratory, SciLifeLabTillämpad fysikMedicinsk bildteknikBiomedicinsk fysik och röntgenfysik
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