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A simple add-on microfluidic appliance for accurately sorting small populations of cells with high fidelity.

ABSTRACT: Current advances in single cell sequencing, gene expression and proteomics require the isolation of single cells, frequently from a very small source population. In this work we describe the design and characterization of a manually operated microfluidic cell sorter that 1) can accurately sort single or small groups of cells from very small cell populations with minimal losses, 2) that is easy to operate and that can be used in any laboratory that has a basic fluorescent microscope and syringe pump, 3) that can be assembled within minutes, 4) that can sort cells in very short time (minutes) with minimum cell stress, 5) that is cheap and reusable. This microfluidic sorter is made from hard plastic material (PMMA) into which microchannels are directly milled with hydraulic diameter of 70 ?m. Inlet and outlet reservoirs are drilled through the chip. Sorting occurs through hydrodynamic switching ensuring low hydrodynamic shear stresses, which were modeled or experimentally confirmed to be below the cell damage threshold. Manually operated, the maximum sorting frequencies were approximately 10 cells per minute. Experiments verified that cell sorting operations could be achieved in as little as 15 minutes, including the assembly and testing of the sorter. In only one out of 10 sorting experiments the sorted cells were contaminated with another cell type. This microfluidic cell sorter represents an important capability for protocols requiring fast isolation of single cells from small number of rare cell populations.

SUBMITTER: Grad M

PROVIDER: S-EPMC3883503 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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A simple add-on microfluidic appliance for accurately sorting small populations of cells with high fidelity.

Grad Michael M Young Erik F EF Smilenov Lubomir L Brenner David J DJ Attinger Daniel D

Journal of micromechanics and microengineering : structures, devices, and systems 20130101 11

Current advances in single cell sequencing, gene expression and proteomics require the isolation of single cells, frequently from a very small source population. In this work we describe the design and characterization of a manually operated microfluidic cell sorter that 1) can accurately sort single or small groups of cells from very small cell populations with minimal losses, 2) that is easy to operate and that can be used in any laboratory that has a basic fluorescent microscope and syringe p ...[more]

PMID: 24409041

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Project description:Magnetic nanoparticles (MNPs) are useful for many biomedical applications, but it is challenging to synthetically produce them in large numbers with uniform properties and surface functionalization. Magnetotactic bacteria (MTB) produce magnetosomes with homogenous sizes, shapes, and magnetic properties. Consequently, there is interest in using MTB as biological factories for MNP production. Nonetheless, MTB can only be grown to low yields, and wild-type strains produce low numbers of MNPs/bacterium. There are also limited technologies to facilitate the selection of MTB with different magnetic contents, such as MTB with compromised and enhanced biomineralization ability. Here, we describe a magnetic microfluidic platform combined with transient cold/alkaline treatment to temporarily reduce the rapid flagellar motion of MTB without compromising their long-term proliferation and biomineralization ability for separating MTB on the basis of their magnetic contents. This strategy enables live MTB to be enriched, which, to the best of our knowledge, has not been achieved with another previously described magnetic microfluidic device that makes use of ferrofluid and heat. Our device also facilitates the high-throughput (25,000 cells/min) separation of wild-type Magnetospirillum gryphiswaldense (MSR-1) from nonmagnetic ΔmamAB MSR-1 mutants with a sensitivity of up to 80% and isolation purity of up to 95%, as confirmed with a gold-standard fluorescent-activated cell sorter (FACS) technique. This offers a 25-fold higher throughput than other previously described magnetic microfluidic platforms (1,000 cells/min). The device can also be used to isolate Magnetospirillum magneticum (AMB-1) mutants with different ranges of magnetosome numbers with efficiencies close to theoretical estimates. We believe this technology will facilitate the magnetic characterization of genetically engineered MTB for a variety of applications, including using MTB for large-scale, controlled MNP production.IMPORTANCE Our magnetic microfluidic technology can greatly facilitate biological applications with magnetotactic bacteria, from selection and screening to analysis. This technology will be of interest to microbiologists, chemists, and bioengineers who are interested in the biomineralization and selection of magnetotactic bacteria (MTB) for applications such as directed evolution and magnetogenetics.

Dataset Information

A simple add-on microfluidic appliance for accurately sorting small populations of cells with high fidelity.

Publications

A simple add-on microfluidic appliance for accurately sorting small populations of cells with high fidelity.

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