vin Ke

Noise and Bandwidth of Current Recordings from Submicrometer Pores and Nanopores

Nanopores and submicrometer pores have recently been explored for applications ranging from detection of single molecules, assemblies of nanoparticles, nucleic acids, occurrence of chemical reactions, and unfolding of proteins. Most of these applications rely on monitoring electrical current through these pores, hence the noise and signal bandwidth of these current recordings are critical for achieving accurate and sensitive measurements. In this report, we present a detailed theoretical and experimental study on the noise and signal bandwidth of current recordings from glass and polyethylene terephthalate (PET) membranes that contain a single submicrometer pore or nanopore. We examined the theoretical signal bandwidth of two different pore geometries, and we measured the signal bandwidth of the electronics used to record the ionic current. We also investigated the theoretical noise generated by the substrate material, the pore, and the electronics used to record the current. Employing a combination of theory and experimental results, we were able to predict the noise in current traces recorded from glass and PET pores with no applied voltage with an error of less than 12% in a range of signal bandwidths from 1 to 40 kHz. In approximately half of all experiments, application of a voltage did not significantly increase the noise. In the other half of experiments, however, application of a voltage resulted in an additional source of noise. For these pores, predictions of the noise were usually still accurate within 35% error at signal bandwidths of at least 10 kHz. The power spectra of this extra noise suggested a 1/f(alpha) origin with best fits to the power spectrum for alpha = 0.4-0.8. This work provides the theoretical background and experimental data for understanding the bandwidth requirements and the main sources of noise in current recordings; it will be useful for minimizing noise and achieving accurate recordings.

The Distribution of Polar Ejection Forces Determines the Amplitude of Chromosome Directional Instability

BACKGROUND Polar ejection forces have often been hypothesized to guide directional instability of mitotic chromosomes, but a direct link has never been established. This has led, in part, to the resurgence of alternative theories. By taking advantage of extremely precise femtosecond pulsed laser microsurgery, we abruptly alter the magnitude of polar ejection forces by severing vertebrate chromosome arms. RESULTS Reduction of polar ejection forces increases the amplitude of directional instability without altering other characteristics, thus establishing a direct link between polar ejection forces and the direction of chromosome movements. We find that polar ejection forces limit the range of chromosome oscillations by increasing the probability that motors at a leading kinetochore abruptly disengage or turn off, leading to a direction reversal. CONCLUSIONS From the relation between the change in oscillation amplitude and the amount a chromosome arm is shortened, we are able to map the distribution of polar ejection forces across the spindle, which is surprisingly different from previously assumed distributions. These results allow us to differentiate between the mechanisms proposed to underlie the directional instability of chromosomes.

Ultrafast laser fabrication of submicrometer pores in borosilicate glass

We demonstrate rapid fabrication of submicrometer-diameter pores in borosilicate glass using femtosecond laser machining and subsequent wet-etch techniques. This approach allows direct and repeatable fabrication of high-quality pores with diameters of 400-800 nm. Such small pores coupled with the desirable electrical and chemical properties of glass enable sensitive resistive-pulse analysis to determine the size and concentration of macromolecules and nanoparticles. Plasma-enhanced chemical vapor deposition allows further reduction of pore diameters to below 300 nm.

Kinetic properties of ASC protein aggregation in epithelial cells.

Apoptosis‐associated speck‐like protein with CARD domain (ASC), an adaptor protein composed of caspase recruitment and pyrin domains, can efficiently self‐associate to form a large spherical structure, called a speck. Although ASC aggregation is generally involved with both inflammatory processes and apoptosis, the detailed dynamics of speck formation have not been characterized. In this report, speck formation in HeLa cells transfected with ASC is examined by time‐lapse live‐imaging by confocal laser scanning microscopy. The results show that ASC aggregation is a very rapid and tightly regulated process. Prior to speck formation, soluble ASC aggregation is a low probability event, and the affinity of ASC subunits for one another is very low. Following a speck nucleation event, the affinity for further addition of ASC subunits increases dramatically, and aggregation is a highly energetically favorable reaction (Gibbs free energy ∼ −40 kJ/mol). This leads to a rapid depletion of soluble ASC, making it highly unlikely that a second speck will form inside the same cell and assuring that speck formation is “all or none,” with a well‐defined end point. Comparison with kinetic models of the aggregation process indicates diffusion, instead of active transport, is the dominant process for speck growth. Though speck formation and aggresome formation share some properties, we show that the two processes are distinct. J. Cell. Physiol. 222: 738–747, 2010.

Power scalable >25 W supercontinuum laser from 2 to 2.5 μm with near-diffraction-limited beam and low output variability

A power scalable thulium-doped fiber-amplifier-based supercontinuum (SC) laser covering the shortwave infrared region from 2 to 2.5 μm is demonstrated. The SC laser has an average power up to 25.7 W and a spectral density of >12 dBm/nm. Power scalability of the laser is proven by showing that the SC laser maintains a nearly constant spectral output, beam quality (M(2) measurements), and output spectral stability as the SC average power is scaled from 5 to 25.7 W average output power. We verify that the SC laser beam is nearly diffraction limited with an M(2)<1.2 for all power levels. Output spectral stability measurements with power scaling show a radiometric variability of <0.8% across the entire SC spectrum.

Mid-infrared absorption spectroscopy and differential damage in vitro between lipids and proteins by an all-fiber-integrated supercontinuum laser

We identify and differentially damage lipids and proteins using wavelengths between 2.6 and 3.8 mum from a fiber-based supercontinuum (SC) laser. Absorption spectroscopy of the constituents of normal artery and atherosclerotic plaque, including adipose tissue, macrophages and foam cells, are measured by a SC laser in the mid-infrared. By using the laser light within the C-H fatty acid and cholesterol esters absorption band, we also demonstrate differential damage of lipid-rich adipose tissue without damaging the protein-rich blood vessel wall. The experiments use a novel SC laser that is all-fiber-integrated with no moving parts, covers a continuous spectrum ranging from approximately 0.8 to beyond 4.2 microm, and outputs a time-averaged power scalable up to 10.5 W.

Photothermolysis of sebaceous glands in human skin ex vivo with a 1,708 nm Raman fiber laser and contact cooling

Wavelengths near ∼1,720 nm are of interest for targeting fat/lipid‐rich tissues due to the high absorption coefficient of human fat and low water scattering and absorption. In this study, a 1,708 nm laser was built and shown to selectively target fat/lipid adjacent to porcine heart and dermis and then used to damage dermal sebaceous glands in human skin.

Field trial of active remote sensing using a high-power short-wave infrared supercontinuum laser

Field trial results of a 5 W all-fiber broadband supercontinuum (SC) laser covering the short-wave infrared (SWIR) wavelength bands from ~1.55 to 2.35 μm are presented. The SC laser is kept on a 12 story tower at the Wright Patterson Air Force Base and propagated through the atmosphere to a target 1.6 km away. Beam quality of the SC laser after propagating through 1.6 km is studied using a SWIR camera and show a near diffraction limited beam with an M(2) value of <1.3. The SC laser is used as the illumination source to perform spectral reflectance measurements of various samples at 1.6 km, and the results are seen to be in good agreement with in-lab measurements using a conventional lamp source. Spectral stability measurements are performed after atmospheric propagation through 1.6 km and show a relative variability of ~4%-8% across the spectrum depending on the atmospheric turbulence effects. Spectral stability measurements are also performed in-lab and show a relative variability of <0.6% across the spectrum.

Mid-IR super-continuum generation

A Mid-InfraRed FIber Laser (MIRFIL) has been developed that generates super-continuum covering the spectral range from 0.8 to 4.5 microns with a time-averaged power as high as 10.5W. The MIRFIL is an all-fiber integrated laser with no moving parts and no mode-locked lasers that uses commercial off-the-shelf parts and leverages the mature telecom/fiber optics platform. The MIRFIL power can be easily scaled by changing the repetition rate and modifying the erbium-doped fiber amplifier. Some of the applications using the super-continuum laser will be described in defense, homeland security and healthcare. For example, the MIRFIL is being applied to a catheter-based medical diagnostic system to detect vulnerable plaque, which is responsible for most heart attacks resulting from hardening-of-the-arteries or atherosclerosis. More generally, the MIRFIL can be a platform for selective ablation of lipids without damaging normal protein or smooth muscle tissue.

Tower testing of a 64W shortwave infrared supercontinuum laser for use as a hyperspectral imaging illuminator

Hyperspectral imaging systems are currently used for numerous activities related to spectral identification of materials. These passive imaging systems rely on naturally reflected/emitted radiation as the source of the signal. Thermal infrared systems measure radiation emitted from objects in the scene. As such, they can operate at both day and night. However, visible through shortwave infrared systems measure solar illumination reflected from objects. As a result, their use is limited to daytime applications. Omni Sciences has produced high powered broadband shortwave infrared super-continuum laser illuminators. A 64-watt breadboard system was recently packaged and tested at Wright-Patterson Air Force Base to gauge beam quality and to serve as a proof-of-concept for potential use as an illuminator for a hyperspectral receiver. The laser illuminator was placed in a tower and directed along a 1.4km slant path to various target materials with reflected radiation measured with both a broadband camera and a hyperspectral imaging system to gauge performance.