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Elasto-inertial microfluidics for bacteria separation from whole blood for sepsis diagnostics
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. mafaridi@kth.se. (Clinical Microfluidics)ORCID iD: 0000-0003-1176-0905
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. (Clinical Microfluidics)ORCID iD: 0000-0001-5199-0663
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. (Clinical Microfluidics)
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
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2017 (English)In: Journal of Nanobiotechnology, ISSN 1477-3155, E-ISSN 1477-3155, Vol. 15, article id 3Article in journal (Refereed) Published
Abstract [en]

Background: Bloodstream infections (BSI) remain a major challenge with high mortality rate, with an incidence that is increasing worldwide. Early treatment with appropriate therapy can reduce BSI-related morbidity and mortality. However, despite recent progress in molecular based assays, complex sample preparation steps have become critical roadblock for a greater expansion of molecular assays. Here, we report a size based, label-free, bacteria separation from whole blood using elasto-inertial microfluidics.

Results: In elasto-inertial microfluidics, the viscoelastic flow enables size based migration of blood cells into a non- Newtonian solution, while smaller bacteria remain in the streamline of the blood sample entrance and can be sepa- rated. We first optimized the flow conditions using particles, and show continuous separation of 5 μm particles from 2 μm at a yield of 95% for 5 μm particle and 93% for 2 μm particles at respective outlets. Next, bacteria were continu- ously separated at an efficiency of 76% from undiluted whole blood sample.

Conclusion: We demonstrate separation of bacteria from undiluted while blood using elasto-inertial microfluidics. The label-free, passive bacteria preparation method has a great potential for downstream phenotypic and molecular analysis of bacteria. 

Place, publisher, year, edition, pages
BioMed Central, 2017. Vol. 15, article id 3
Keywords [en]
Micro particle separation, Elasto-inertial microfluidics, Sepsis, Sample preparation
National Category
Medical Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-200300DOI: 10.1186/s12951-016-0235-4ISI: 000391073000001Scopus ID: 2-s2.0-85008198016OAI: oai:DiVA.org:kth-200300DiVA, id: diva2:1068014
Projects
RAPP_ID
Funder
EU, European Research Council, 115153
Note

QC 20170124

Available from: 2017-01-24 Created: 2017-01-24 Last updated: 2019-02-28Bibliographically approved
In thesis
1. Bioparticle Manipulation using Acoustophoresis and Inertial Microfluidics
Open this publication in new window or tab >>Bioparticle Manipulation using Acoustophoresis and Inertial Microfluidics
2017 (English)Doctoral thesis, comprehensive summary (Other academic)
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.

Place, publisher, year, edition, pages
Stockholm, Sweden: Kungliga Tekniska högskolan, 2017. p. 68
Series
TRITA-BIO-Report, ISSN 1654-2312 ; 2017:4
National Category
Medical Biotechnology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-200304 (URN)978-91-7729-264-7 (ISBN)
Public defence
2017-02-16, Gard-Aulan, Nobels vägen 18, Solna, Stockholm, 10:00 (English)
Opponent
Supervisors
Note

QC 20170124

Available from: 2017-01-24 Created: 2017-01-24 Last updated: 2017-01-24Bibliographically approved
2. Point of care microfluidic tool development for resource limited settings
Open this publication in new window or tab >>Point of care microfluidic tool development for resource limited settings
2019 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The development of point of care diagnostics using recent advances in microfluidics have the potential to transform health care in several ways, especially in resource limited settings with limited access to advanced health care infrastructure. However, translating a point of care device to reality is often a challenging task because of the complexities involved in integrating a number of diverse engineering concepts into an easy to use, accurate and portable device. This thesis focuses on miniaturization of crucial diagnostic laboratory tools, that can be used in a portable point of care format without compromising on the accuracy or performance. The first part of the thesis (Paper I-III) focuses on understanding and applying elasto-inertial microfluidics, which is a label-free and passive bio-particle sorting and separation method. A basic understanding of particle trajectories in both inertial (Paper I) and visco-elastic flows (Paper II) is established, followed by an investigation on the combined effects of inertia and elasticity (Paper III). The second part of the thesis (Paper IV-VI) focuses on developing integrated microfluidic platforms, each of which addresses different aspects of point of care diagnostic applications. The applications include neonatal diagnostics using a hand-driven Slipdisc technique (Paper IV), rapid nucleic acid quantification using a novel precipitate-based detection on a centrifugal microfluidics platform (Paper V), and hematocrit level measurement in blood using a portable lab-on- Disc platform operated by a mobile phone (Paper VI). The proof of concept microfluidic tools presented in the scope of this thesis have the potential to replace a number of functions of standard laboratory equipment, at a fraction of the price and without compromising performance. Hence, the different methods developed should contribute towards decentralization of medical testing laboratories, making healthcare accessible to one and all.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2019. p. 63
Series
TRITA-CBH-FOU ; 2019:15
Keywords
Blood, control, cell separation, centrifugal microfluidics, diagnostics, elasto-inertial, hematocrit level, microfluidics, neonatal diagnostics, nucleic acid quantification, point of care, particle focusing, resource limited settings.
National Category
Medical and Health Sciences Engineering and Technology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-244825 (URN)978-91-7873-122-0 (ISBN)
Public defence
2019-03-29, Air & Fire auditorium, Science for Life Laboratory, Tomtebodavägen 23, Solna, 10:00 (English)
Opponent
Supervisors
Note

QC 20190228

Available from: 2019-02-28 Created: 2019-02-28 Last updated: 2019-02-28Bibliographically approved

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Faridi, Muhammad AsimRamachandraiah, HarishaRussom, Aman

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