R. Sooryakumar

Magnetic Tweezers‐Based 3D Microchannel Electroporation for High‐Throughput Gene Transfection in Living Cells

A novel high-throughput magnetic tweezers-based 3D microchannel electroporation system capable of transfecting 40 000 cells/cm(2) on a single chip for gene therapy, regenerative medicine, and intracellular detection of target mRNA for screening cellular heterogeneity is reported. A single cell or an ordered array of individual cells are remotely guided by programmable magnetic fields to poration sites with high (>90%) cell alignment efficiency to enable various transfection reagents to be delivered simultaneously into the cells. The present technique, in contrast to the conventional vacuum-based approach, is significantly gentler on the cellular membrane yielding >90% cell viability and, moreover, allows transfected cells to be transported for further analysis. Illustrating the versatility of the system, the GATA2 molecular beacon is delivered into leukemia cells to detect the regulation level of the GATA2 gene that is associated with the initiation of leukemia. The uniform delivery and a sharp contrast of fluorescence intensity between GATA2 positive and negative cells demonstrate key aspects of the platform for gene transfer, screening and detection of targeted intracellular markers in living cells.

On-chip magnetic separation and encapsulation of cells in droplets

Single cell study is gaining importance because of the cell-to-cell variation that exists within cell population, even after significant initial sorting. Analysis of such variation at the gene expression level could impact single cell functional genomics, cancer, stem-cell research, and drug screening. The on-chip monitoring of individual cells in an isolated environment would prevent cross-contamination, provide high recovery yield, and enable study of biological traits at a single cell level. These advantages of on-chip biological experiments is a significant improvement for a myriad of cell analyses methods, compared to conventional methods, which require bulk samples and provide only averaged information on cell structure and function. We report on a device that integrates a mobile magnetic trap array with microfluidic technology to provide the possibility of separation of immunomagnetically labeled cells and their encapsulation with reagents into picoliter droplets for single cell analysis. The simultaneous reagent delivery and compartmentalization of the cells immediately following sorting are all performed seamlessly within the same chip. These steps offer unique advantages such as the ability to capture cell traits as originated from its native environment, reduced chance of contamination, minimal use of the reagents, and tunable encapsulation characteristics independent of the input flow. Preliminary assay on cell viability demonstrates the potential for the device to be integrated with other up- or downstream on-chip modules to become a powerful single-cell analysis tool.

Mechanical properties of high porosity low-k dielectric nano-films determined by Brillouin light scattering

Integrating nanometre sized pores into hybrid organic-inorganic interconnect layers is one of the key approaches being undertaken by the semiconductor industry to sustain the continued scale down of micro-electronic devices. While increasing porosity of the layers achieves the desirable lowering of the dielectric constant (k), it also has the potential to reduce mechanical and thermal stability and degrade device functionality. We report on Brillouin light scattering to measure the independent elastic constants, and thus the mechanical properties, of ultrathin dielectric films with porosity levels up to 45%, the highest in the industry. Longitudinal and transverse acoustic standing mode type excitations were observed from sub 200nm thick low-k thin films, and their frequency dispersion and associated light scattering intensities were utilized to determine Poisson’s ratio (ν)and Young’s modulus (E). In comparison with SiO2 and non-porous low-k materials, significant modifications were found in ν and E of these highly porous carbon-doped SiO2 (Si‐O‐C‐H) and amorphous carbon (a-C:H) low-k interlayer dielectrics.

Elastic properties of porous low-k dielectric nano-films

Low-k dielectrics have predominantly replaced silicon dioxide as the interlayer dielectric for interconnects in state of the art integrated circuits. In order to further reduce interconnect RC delays, additional reductions in k for these low-k materials are being pursued via the introduction of controlled levels of porosity. The main challenge for such dielectrics is the substantial reduction in elastic properties that accompanies the increased pore volume. We report on Brillouin light scattering measurements used to determine the elastic properties of these films at thicknesses well below 200 nm, which are pertinent to their introduction into present ultralarge scale integrated technology. The observation of longitudinal and transverse standing wave acoustic resonances and their transformation into traveling waves with finite in-plane wave vectors provides for a direct non-destructive measure of the principal elastic constants that characterize the elastic properties of these porous nano-scale films. The...

Manipulation of Magnetically Labeled and Unlabeled Cells with Mobile Magnetic Traps

A platform of discrete microscopic magnetic elements patterned on a surface offers dynamic control over the motion of fluid-borne cells by reprogramming the magnetization within the magnetic bits. T-lymphocyte cells tethered to magnetic microspheres and untethered leukemia cells are remotely manipulated and guided along desired trajectories on a silicon surface by directed forces with average speeds up to 20 microm/s. In addition to navigating cells, the microspheres can be operated from a distance to push biological and inert entities and act as local probes in fluidic environments.

Elastic constants of face-centered-cubic cobalt

The wave vector dependence of Rayleigh and higher order Sezawa elastic waves in single crystalline epitaxial face-centered-cubic (fcc) Co layers are measured by Brillouin light scattering at room temperature. The dispersion of the mode velocities allows the independent elastic constants to be determined for this cubic phase of cobalt. These results compare very favorably to previous determinations of the elastic constants measured above 700 K from the high-temperature fcc phase of pure Co. Deviations from theoretical estimates are discussed.

Ultra-localized single cell electroporation using silicon nanowires

Analysis of cell-to-cell variation can further the understanding of intracellular processes and the role of individual cell function within a larger cell population. The ability to precisely lyse single cells can be used to release cellular components to resolve cellular heterogeneity that might be obscured when whole populations are examined. We report a method to position and lyse individual cells on silicon nanowire and nanoribbon biological field effect transistors. In this study, HT-29 cancer cells were positioned on top of transistors by manipulating magnetic beads using external magnetic fields. Ultra-rapid cell lysis was subsequently performed by applying 600-900 mV(pp) at 10 MHz for as little as 2 ms across the transistor channel and the bulk substrate. We show that the fringing electric field at the device surface disrupts the cell membrane, leading to lysis from irreversible electroporation. This methodology allows rapid and simple single cell lysis and analysis with potential applications in medical diagnostics, proteome analysis and developmental biology studies.

Light-Scattering Study of the Normal Human Eye Lens: Elastic Properties and Age Dependence

The human ocular lens is a tissue capable of changing its shape to dynamically adjust the optical power of the eye, a function known as accommodation, which gradually declines with age. This capability is the response of the lens tissue to external forces, which, in turn, is modulated by the biomechanical characteristics of lens tissues. In order to investigate the contributions of lens sclerosis to loss of accommodation, we report on in vitro confocal Brillouin light scattering studies of human ocular lenses spanning over a 30-70 year age range. Using this nondestructive measurement method, we determined that the longitudinal bulk modulus (average ± SD) of the lens nucleus (2.79 ± 0.14 GPa) was consistently greater than the bulk modulus of the lens cortex (2.36 ± 0.09 GPa). Moreover, our results showed that these differences were not age dependent over the 40 year age range that we evaluated using healthy lens tissues. Our results are consistent with the hypothesis that an age-dependent change in the bulk modulus of lens tissues does not fully account for the natural decline of accommodation.

Brillouin light scattering studies of the mechanical properties of ultrathin low-k dielectric films

In an effort to reduce RC time delays that accompany decreasing feature sizes, low-k dielectric films are rapidly emerging as potential replacements for silicon dioxide (SiO2) at the interconnect level in integrated circuits. The main challenge in low-k materials is their substantially weaker mechanical properties that accompany the increasing pore volume content needed to reduce k. We show that Brillouin light scattering is an excellent nondestructive technique to monitor and characterize the mechanical properties of these porous films at thicknesses well below 200nm that are pertinent to present applications. Observation of longitudinal and transverse standing wave acoustic resonances and the dispersion that accompany their transformation into traveling waves with finite in-plane wave vectors provides for a direct measure of the principal elastic constants that completely characterize the mechanical properties of these ultrathin films. The mode amplitudes of the standing waves, their variation within th...

Dispersion of elastic waves in supported CaF2 films

The velocities of the principal and several higher‐order elastic modes localized in CaF2 films grown epitaxially on Si(111) have been measured using Brillouin scattering. The effective elastic constants of the epilayers are evaluated from a fit to the mode dispersion. With decreasing film thicknesses from about 1500 A reductions from the bulk elastic constants are found. Thicker films are well characterized by bulk parameters.