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Intuitive, image-based cell sorting using optofluidic cell sorting.

ABSTRACT: We present a microfluidic cell-sorting device which augments microscopy with the capability to perform facile image-based cell sorting. This combination enables intuitive, complex phenotype sorting based on spatio-temporal fluorescence or cell morphology. The microfluidic device contains a microwell array that can be passively loaded with mammalian cells via sedimentation and can be subsequently inspected with microscopy. After inspection, we use the scattering force from a focused infrared laser to levitate cells of interest from their wells into a flow field for collection. First, we demonstrate image-based sorting predicated on whole-cell fluorescence, which could enable sorting based on temporal whole-cell fluorescence behavior. Second, we demonstrate image-based sorting predicated on fluorescence localization (nuclear vs whole-cell fluorescence), highlighting the capability of our approach to sort based on imaged subcellular events, such as localized protein expression or translocation events. We achieve postsort purities up to 89% and up to 155-fold enrichment of target cells. Optical manipulation literature and a direct cell viability assay suggest that cells remain viable after using our technique. The architecture is highly scalable and supports over 10 000 individually addressable trap sites. Our approach enables sorting of significant populations based on subcellular spatio-temporal information, which is difficult or impossible with existing widespread sorting technologies.

SUBMITTER: Kovac JR

PROVIDER: S-EPMC2536524 | biostudies-literature | 2007 Dec

REPOSITORIES: biostudies-literature

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Intuitive, image-based cell sorting using optofluidic cell sorting.

Kovac J R JR Voldman J J

Analytical chemistry 20071116 24

We present a microfluidic cell-sorting device which augments microscopy with the capability to perform facile image-based cell sorting. This combination enables intuitive, complex phenotype sorting based on spatio-temporal fluorescence or cell morphology. The microfluidic device contains a microwell array that can be passively loaded with mammalian cells via sedimentation and can be subsequently inspected with microscopy. After inspection, we use the scattering force from a focused infrared lase ...[more]

PMID: 18004819

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Project description:This work proposes a novel design composed of graphene nanoribbons-based optofluidic tweezers to manipulate and sort bio-particles with radii below 2.5 nm. The suggested structure has been numerically investigated by the finite difference time domain (FDTD) method employing Maxwell's stress tensor analysis (MST). The finite element method (FEM) has been used to obtain the electrostatic response of the proposed structure. The tweezer main path is a primary channel in the center of the structure, where the microfluidic flow translates the nanoparticle toward this channel. Concerning the microfluid's drag force, the nanoparticles tend to move along the length of the main channel. The graphene nanoribbons are fixed near the main channel at different distances to exert optical forces on the moving nanoparticles in the perpendicular direction. In this regard, sub-channels embedding in the hBN layer on the Si substrate deviate bio-particles from the main path for particular nanoparticle sizes and indices. Intense hotspots with electric field enhancements up to 900 times larger than the incident light are realized inside and around the graphene ribbons. Adjusting the gap distance between the graphene nanoribbon and the main channel allows us to separate the individual particle with a specific size from others, thus guiding that in the desired sub-channel. Furthermore, we demonstrated that in a structure with a large gap between channels, particles experience weak field intensity, leading to a low optical force that is insufficient to detect, trap, and manipulate nanoparticles. By varying the chemical potential of graphene associated with the electric field intensity variations in the graphene ribbons, we realized tunability in sorting nanoparticles while structural parameters remained constant. In fact, by adjusting the graphene Fermi level via the applied gate voltage, nanoparticles with any desired radius will be quickly sorted. Moreover, we exhibited that the proposed structure could sort nanoparticles based on their refractive indices. Therefore, the given optofluidic tweezer can easily detect bio-particles, such as cancer cells and viruses of tiny size.

Dataset Information

Intuitive, image-based cell sorting using optofluidic cell sorting.

Publications

Intuitive, image-based cell sorting using optofluidic cell sorting.

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