Density-Gradient Mediated Band Extraction of Leukocytes from Whole Blood Using Centrifugo-Pneumatic Siphon Valving on Centrifugal Microfluidic Discs.

Authors:
David J Kinahan
David J Kinahan
Dublin City University
Ireland
Niamh A Kilcawley
Niamh A Kilcawley
Biomedical Diagnostics Institute
Philip L Early
Philip L Early
School of Physical Sciences
Agra | India
Macdara T Glynn
Macdara T Glynn
Biomedical Diagnostics Institute
Dr. Jens Ducree, Dr. rer. nat. habil. Dipl. Phys.
Dr. Jens Ducree, Dr. rer. nat. habil. Dipl. Phys.
Fraunhofer Project Centre at Dublin City University
Professor (Full)
microfluidics, Lab-on-a-Chip, hydrodynanmics, business development, project management, organisational leadership
Glasnevin, Dublin 9 | Ireland

PLoS One 2016 11;11(5):e0155545. Epub 2016 May 11.

School of Physical Sciences, Dublin City University, Glasnevin, Dublin 9, Ireland.

Here we present retrieval of Peripheral Blood Mononuclear Cells by density-gradient medium based centrifugation for subsequent analysis of the leukocytes on an integrated microfluidic "Lab-on-a-Disc" cartridge. Isolation of white blood cells constitutes a critical sample preparation step for many bioassays. Centrifugo-pneumatic siphon valves are particularly suited for blood processing as they function without need of surface treatment and are 'low-pass', i.e., holding at high centrifugation speeds and opening upon reduction of the spin rate. Both 'hydrostatically' and 'hydrodynamically' triggered centrifugo-pneumatic siphon valving schemes are presented. Firstly, the geometry of the pneumatic chamber of hydrostatically primed centrifugo-pneumatic siphon valves is optimised to enable smooth and uniform layering of blood on top of the density-gradient medium; this feature proves to be key for efficient Peripheral Blood Mononuclear Cell extraction. A theoretical analysis of hydrostatically primed valves is also presented which determines the optimum priming pressure for the individual valves. Next, 'dual siphon' configurations for both hydrostatically and hydrodynamically primed centrifugo-pneumatic siphon valves are introduced; here plasma and Peripheral Blood Mononuclear Cells are extracted through a distinct siphon valve. This work represents a first step towards enabling on disc multi-parameter analysis. Finally, the efficiency of Peripheral Blood Mononuclear Cells extraction in these structures is characterised using a simplified design. A microfluidic mechanism, which we termed phase switching, is identified which affects the efficiency of Peripheral Blood Mononuclear Cell extraction.

Download full-text PDF

Source
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0155545PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4864222PMC

Still can't find the full text of the article?

Sign up to send a request to the authors directly.
July 2017
5 Reads
5 PubMed Central Citations(source)
3.23 Impact Factor

Publication Analysis

Top Keywords

centrifugo-pneumatic siphon
20
blood mononuclear
20
peripheral blood
20
siphon valves
12
mononuclear cells
12
blood
9
cell extraction
8
mononuclear cell
8
density-gradient medium
8
siphon valving
8
primed centrifugo-pneumatic
8
efficiency peripheral
8
hydrostatically primed
8
siphon
6
peripheral
5
centrifugo-pneumatic
5
valves
5
mononuclear
5
chamber hydrostatically
4
valves optimised
4

Altmetric Statistics

References

(Supplied by CrossRef)
The centrifugal microfluidic Bio-Disk platform
J Ducrée et al.
Journal of Micromechanics and Microengineering 2007
Lab on a CD
M Madou et al.
Annu Rev Biomed Eng 2006
Centrifugal microfluidic platforms: advanced unit operations and applications
O Strohmeier et al.
Chemical Society Reviews 2015
A lab-on-CD prototype for high-speed blood separation
J Zhang et al.
Journal of micromechanics and microengineering 2008
Geometry effects on blood separation rate on a rotating disc
T-H Kim et al.
Sensors and Actuators B: Chemical 2013

Similar Publications

Identification of a new turncurtovirus in the leafhopper Circulifer haematoceps and the host plant species Sesamum indicum.

Virus Genes 2018 Oct 11. Epub 2018 Oct 11.

The Biodesign Center of Fundamental and Applied Microbiomics, School of Life Sciences, Center for Evolution and Medicine, Arizona State University, 1001 S. McAllister Ave, Tempe, AZ, 85287-5001, USA.

Turncurtoviruses (family: Geminiviridae; genus: Turncurtovirus) appear to have a high degree of genetic variation in Iran. Leafhoppers of the species Circulifer haematoceps (Mulsant and Rey, 1855) (family: Cicadellidae) were collected in 2014 from three geographical regions in south-eastern Iran (Orzoeyeh, Jiroft and Sirjan; Kerman province) and screened for the presence of turncurtoviruses using a combination of PCR and rolling circle amplification (RCA) methods. Eleven genomes of turncurtovirus were recovered and sequenced. Read More

View Article
October 2018
40 Reads
1.84 Impact Factor

Human neuroblastoma SH-SY5Y cells treated with okadaic acid express phosphorylated high molecular weight tau-immunoreactive protein species.

J Neurosci Methods 2018 Sep 29. Epub 2018 Sep 29.

Department of Neuroscience, Croatian Institute for Brain Research, University of Zagreb Medical School, Zagreb, Croatia. Electronic address:

Background: Early stages of Alzheimer's disease (AD) are characterized by high phosphorylation of microtubule-associated protein tau, which may result from the downregulation of protein phosphatases.

New Method: In order to model phosphatase downregulation and analyze its effect on tau aggregation in vitro, we treated neuroblastoma SH-SY5Y cells with okadaic acid (OA), a protein phosphatase inhibitor, and examined high molecular weight phospho-tau species.

Results And Comparison With Existing Methods: OA treatment led to the appearance of heat-stable protein species with apparent molecular weight around 100 kDa, which were immunoreactive to anti-tau antibodies against phosphorylated Ser202 and Ser396. Read More

View Article
September 2018
12 Reads
2.02 Impact Factor

MicroRNAs in regulatory T cells.

Cancer Lett 2018 Jun 11;423:80-85. Epub 2018 Mar 11.

Institute of Microbiology, Chinese Academy of Sciences, Beijing, China; Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 101408, China. Electronic address:

View Article
June 2018
7 Reads
5.62 Impact Factor

Co-infection with Toxoplasma gondii and Clostridium perfringens in a postpartum woman with uterine gas gangrene: a case report.

J Obstet Gynaecol Res 2012 Jul 9;38(7):1024-7. Epub 2012 Apr 9.

Department of Obstetrics and Gynecology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.

Toxoplasmosis is a protozoan infection caused by Toxoplasma gondii. We report a case of Toxoplasma gondii and Clostridium perfringens co-infection complicating uterine gas gangrene following a term pregnancy. The histological examination of the necrotic uterine tissues and uterine swab cultures obtained at laparotomy revealed T. Read More

View Article
July 2012
32 Reads
2 PubMed Central Citations(source)
0.93 Impact Factor

Fe/Al synergy in Fe(2)O(3) nanoparticles supported on porous aluminosilicate materials: excelling activities in oxidation reactions.

Chem Commun (Camb) 2010 Nov 9;46(41):7825-7. Epub 2010 Sep 9.

Dpto.Química Orgánica, Universidad de Córdoba, Edificio Marie Curie, Ctra Nnal IV, Km 396, E14014, Córdoba, Spain.

A synergetic Fe-Al effect in Fe(2)O(3) nanoparticles supported on mesoporous aluminosilicates compared to pure siliceous silicates has been demonstrated, for the first time, by a remarkably superior catalytic activity of the former in the microwave-assisted selective oxidation of benzyl alcohol to benzaldehyde. This significant finding, that also deeply influences the acidity of the materials (increasing total and particularly Lewis acidity), can have important consequences in the improved efficiency of these systems in related oxidations as well as in acid catalysed processes. Read More

View Article
November 2010
14 Reads
2 PubMed Central Citations(source)
6.83 Impact Factor