PubMed Health⌕ Search

Biomedical subjects

Nicole Pamme

Publications and source records attributed to Nicole Pamme.

5 recordsLinked to original sources

Continuous sorting of magnetic cells via on-chip free-flow magnetophoresis.

The ability to separate living cells is an essential aspect of cell research. Magnetic cell separation methods are among some of the most efficient methods for bulk cell separation. With the development of microfluidic platforms within the biotechnology sector, the design of miniaturised magnetic cell sorters is desirable. Here, we report the continuous sorting of cells loaded with magnetic nanoparticles in a microfluidic magnetic separation device. Cells were passed through a microfluidic chamber and were deflected from the direction of flow by means of a magnetic field. Two types of cells were studied, mouse macrophages and human ovarian cancer cells (HeLa cells). The deflection was dependent on the magnetic moment and size of the cells as well as on the applied flow rate. The experimentally observed deflection matched well with calculations. Furthermore, the separation of magnetic and non-magnetic cells was demonstrated using the same microfluidic device.

Animals↗

Magnetism and microfluidics.

Magnetic forces are now being utilised in an amazing variety of microfluidic applications. Magnetohydrodynamic flow has been applied to the pumping of fluids through microchannels. Magnetic materials such as ferrofluids or magnetically doped PDMS have been used as valves. Magnetic microparticles have been employed for mixing of fluid streams. Magnetic particles have also been used as solid supports for bioreactions in microchannels. Trapping and transport of single cells are being investigated and recently, advances have been made towards the detection of magnetic material on-chip. The aim of this review is to introduce and discuss the various developments within the field of magnetism and microfluidics.

Biological Assay↗

On-chip free-flow magnetophoresis: continuous flow separation of magnetic particles and agglomerates.

The separation of magnetic microparticles was achieved by on-chip free-flow magnetophoresis. In continuous flow, magnetic particles were deflected from the direction of laminar flow by a perpendicular magnetic field depending on their magnetic susceptibility and size and on the flow rate. Magnetic particles could thus be separated from each other and from nonmagnetic materials. Magnetic and nonmagnetic particles were introduced into a microfluidic separation chamber, and their deflection was studied under the microscope. The magnetic particles were 2.0 and 4.5 microm in diameter with magnetic susceptibilities of 1.12 x 10(-4) and 1.6 x 10(-4) m(3) kg(-1), respectively. The 4.5-microm particles with the larger susceptibility were deflected further from the direction of laminar flow than the 2.0-microm magnetic particles. Nonmagnetic 6-microm polystyrene beads, however, were not deflected at all. Furthermore, agglomerates of magnetic particles were found to be deflected to a larger extent than single magnetic particles. The applied flow rate and the strength and gradient of the applied magnetic field were the key parameters in controlling the deflection. This separation method has a wide applicability since magnetic particles are commonly used in bioanalysis as a solid support material for antigens, antibodies, DNA, and even cells. Free-flow magnetophoretic separations could be hyphenated with other microfluidic devices for reaction and analysis steps to form a micro total analysis system.

Antibodies↗

Counting and sizing of particles and particle agglomerates in a microfluidic device using laser light scattering: application to a particle-enhanced immunoassay.

A microfluidic device for counting and sizing particles and particle agglomerates based on laser light scattering is demonstrated. The particles were confined hydrodynamically and passed through a focused laser beam. Scattering at two different angles, 15 degree and 45 degree, was detected. At an acquisition rate of 10 kHz, a throughput of 150 particles s(-1) was achieved. Scattering intensity was found to depend on particle volume for 2 to 9 microm diameter particles. Size discrimination of particles with a diameter ratio of 1: 2 was accomplished. In addition, the scattering signals of particle agglomerates formed in a particle-enhanced immunoassay for C-reactive protein (CRP) were measured. Scattering intensity was found to be dependent on the CRP concentration, 100 ng CRP per mL could be detected. The particle counting method presented is generic and can be employed in a wide variety of assays as well as for cell counting and particle counting.

C-Reactive Protein↗

Analysis of polynitrophenols and hexyl by liquid chromatography-mass spectrometry using atmospheric pressure ionisation methods and a volatile ion-pairing reagent.

An LC-MS method for the determination of picric acid (2,4,6-trinitrophenol), its reductive transformation products picramic acid (2-amino-4,6-dinitrophenol) and iso-picramic acid (4-amino-2,6-dinitrophenol) and hexyl (2,2',4,4',6,6'-hexanitrodiphenylamine) has been developed. The analytes were separated using ion-pairing chromatography with a volatile ion-pairing reagent suitable for subsequent MS detection. The performance of an atmospheric pressure chemical ionisation (APCI) and an electrospray ionisation (ESI) interface was compared. ESI-MS is more sensitive for the analytes, especially for hexyl and picric acid, APCI-MS delivered more fragments necessary for unequivocal identification. With LC-ESI-MS limits of detection using single ion monitoring (SIM) mode are 4 ng (iso-picramic acid), 800 pg (picramic acid), 400 pg (picric acid) and 80 pg (hexyl). For quantification, 15N-picric acid was used as an internal standard. Using this new method, the degradation of picric acid in soil was monitored in a laboratory study. Furthermore, the presence of picramic acid was for the first time verified in soil samples from a former ammunition plant.

Atmospheric Pressure↗