Noninvasive in vivo imaging reveals differences between tectorial membrane and basilar membrane traveling waves in the mouse cochlea.

Sound is encoded within the auditory portion of the inner ear, the cochlea, after propagating down its length as a traveling wave. For over half a century, vibratory measurements to study cochlear traveling waves have been made using invasive approaches such as laser Doppler vibrometry. Although these studies have provided critical information regarding the nonlinear processes within the living cochlea that increase the amplitude of vibration and sharpen frequency tuning, the data have typically been limited to point measurements of basilar membrane vibration. In addition, opening the cochlea may alter its function and affect the findings. Here we describe volumetric optical coherence tomography vibrometry, a technique that overcomes these limitations by providing depth-resolved displacement measurements at 200 kHz inside a 3D volume of tissue with picometer sensitivity. We studied the mouse cochlea by imaging noninvasively through the surrounding bone to measure sound-induced vibrations of the sensory structures in vivo, and report, to our knowledge, the first measures of tectorial membrane vibration within the unopened cochlea. We found that the tectorial membrane sustains traveling wave propagation. Compared with basilar membrane traveling waves, tectorial membrane traveling waves have larger dynamic ranges, sharper frequency tuning, and apically shifted positions of peak vibration. These findings explain discrepancies between previously published basilar membrane vibration and auditory nerve single unit data. Because the tectorial membrane directly overlies the inner hair cell stereociliary bundles, these data provide the most accurate characterization of the stimulus shaping the afferent auditory response available to date.

Noncontact orientation of objects in three-dimensional space using magnetic levitation.

This paper describes several noncontact methods of orienting objects in 3D space using Magnetic Levitation (MagLev). The methods use two permanent magnets arranged coaxially with like poles facing and a container containing a paramagnetic liquid in which the objects are suspended. Absent external forcing, objects levitating in the device adopt predictable static orientations; the orientation depends on the shape and distribution of mass within the objects. The orientation of objects of uniform density in the MagLev device shows a sharp geometry-dependent transition: an analytical theory rationalizes this transition and predicts the orientation of objects in the MagLev device. Manipulation of the orientation of the levitating objects in space is achieved in two ways: (i) by rotating and/or translating the MagLev device while the objects are suspended in the paramagnetic solution between the magnets; (ii) by moving a small external magnet close to the levitating objects while keeping the device stationary. Unlike mechanical agitation or robotic selection, orienting using MagLev is possible for objects having a range of different physical characteristics (e.g., different shapes, sizes, and mechanical properties from hard polymers to gels and fluids). MagLev thus has the potential to be useful for sorting and positioning components in 3D space, orienting objects for assembly, constructing noncontact devices, and assembling objects composed of soft materials such as hydrogels, elastomers, and jammed granular media.

Polarization-sensitive interleaved optical coherence tomography.

We introduce a new strategy for single-mode fiber based polarization-sensitive (PS-) optical coherence tomography (OCT) using orthogonally polarized optical frequency combs (OFC) in the sample arm. The two OFCs are tuned to be interleaved in the spectral domain, permitting simultaneous measurement of both polarization states from the same spatial region C close to the location of zero pathlength delay. The two polarization states of the beam in the sample arm are demultiplexed by interpolation after performing wavelength stabilization via a two-mirror calibration method. The system uses Jones matrix methods to measure quantitatively the round-trip phase retardation B-scans in the sample. A glass plate and quarter-wave plate were measured to validate the accuracy of the birefringence measurement. Further, we demonstrated the potential of this system for biomedical applications by measurement of chicken breast muscle.

Optical separation of heterogeneous size distributions of microparticles on silicon nitride strip waveguides.

We demonstrate two complementary optical separation techniques of dielectric particles on the surface of silicon nitride waveguides. Glass particles ranging from 2 µm to 10 µm in diameter are separated at guided powers below 40 mW. The effects of optical, viscous, and frictional forces on the particles are modeled and experimentally shown to enable separation. Particle interactions are investigated and shown to decrease measured particle velocity without interfering with the overall particle separation distribution. The demonstrated separation techniques have the potential to be integrated with microfluidic structures for cell sorting.

Rapid scanning catheterscope for expanded forward-view volumetric imaging with optical coherence tomography.

We demonstrate a novel catheterscope, based on scanning fiber endoscopy, for volumetric imaging with optical coherence tomography (OCT), which possesses a high resonance frequency (>2 kHz) and a small outer diameter (OD) (1.07 mm). Our design is the fastest volumetric-scanning, forward-viewing catheterscope for OCT, and the scanning package has the smallest OD of any such OCT package published to date. Using a proof-of-operation catheterscope with commercial lenses, we demonstrate high-quality in vivo and ex vivo volumetric imaging and extend the 1.1 mm diameter field of view more than 200-fold by mosaicking. Due to its small OD, short rigid tip length, and fast scan rate, this scope is the leading candidate design to enable early detection and staging of bladder cancer during flexible white light cystoscopy.

Wide-field, full-field optical coherence microscopy for high-axial-resolution phase and amplitude imaging.

An original single-objective, full-field optical coherence microscopy system is reported that is capable of imaging both the phase and the amplitude of semi-transparent samples over a field of view of 17.5 mm×17.5 mm with an axial sectioning resolution of 1.5 µm. A special stack acquisition arrangement ensures optimal reachable imaging depth. Several phase-shifting interferometry algorithms for phase measurement with broadband light are compared theoretically and experimentally. Using the phase information, noninvasive depth-resolved topographic images of multilayer samples are produced to characterize each layer by measuring their defects and curvature with a nanometric scale precision. Using the amplitude information, tomographic images with a constant detection sensitivity of ∼80 dB through the entire field of view are obtained and applied to biological specimens.

Quantitative measurements of strain and birefringence with common-path polarization-sensitive optical coherence tomography.

We demonstrate the first system for optical coherence tomography (OCT) that enables simultaneous measurement of quantitative birefringence and strain in biological samples using a common-path configuration. Owing to its superior phase stability, common-path polarization sensitive optical coherence tomography (CoPPSe-OCT) achieves a sub-nanometer displacement sensitivity of 0.52 nm at an SNR of 48 dB. We utilize CoPPSe-OCT to measure reflectance, birefringence, and strain for distinguishing burnt regions in a birefringent biological sample (chicken breast muscle).

Interleaved optical coherence tomography.

We present a novel and cost-effective technique--interleaved optical coherence tomography (iOCT)--to enhance the imaging speed of swept source OCT systems by acquiring data from multiple lateral positions simultaneously during a single wavelength sweep, using a single detector and a virtually imaged phase array (VIPA) as a multi-band demultiplexer. This technique uses spectral encoding to convert coherence length into higher imaging speed; the speed enhancement factor is independent of the source speed or center wavelength, and the effective A-scan rate scales linearly with sweep speed. The optical configuration requires only a change in the sample arm of a traditional OCT system and preserves the axial resolution and fall-off characteristic of a traditional SS-OCT using the same light source. Using 10 kHz, 20 kHz and 100 kHz sources we provide a first demonstration of image speed enhancement factors of up to 12, 6 and 10, respectively, which yield effective A-scan rates of 120 kHz, 120 kHz and 1 MHz for B-scan imaging, with a sensitivity of up to 82.5 dB. We also show that iOCT can image faster dynamics than traditional OCT B-scan imaging and is capable of 3D biological imaging. The iOCT concept suggests a new route to high-speed OCT imaging for laser developers: that is, by focusing on improving the coherence length and linewidth of existing and emerging sources. Hence, iOCT is a nice complement to ongoing research and commercial efforts to enable faster imaging through development of lasers with faster sweep rates, and offers new hope for existing sources with slow sweep rates and potential for enhancement of coherence length to compete with faster sources to achieve high-speed OCT.

The effects of reduced bit depth on optical coherence tomography phase data.

Past studies of the effects of bit depth on OCT magnitude data concluded that 8 bits of digitizer resolution provided nearly the same image quality as a 14-bit digitizer. However, such studies did not assess the effects of bit depth on the accuracy of phase data. In this work, we show that the effects of bit depth on phase data and magnitude data can differ significantly. This finding has an important impact on the design of phase-resolved OCT systems, such as those measuring motion and the birefringence of samples, particularly as one begins to consider the tradeoff between bit depth and digitizer speed.

Infochemistry: encoding information as optical pulses using droplets in a microfluidic device.

This article describes a new procedure for generating and transmitting a message--a sequence of optical pulses--by aligning a mask (an opaque sheet containing transparent "windows") below a microfluidic channel in which flows an opaque continuous fluid containing transparent droplets. The optical mask encodes the message as a unique sequence of windows that can transmit or block light; the flow of transparent droplets in the channel converts this message into a sequence of optical pulses. The properties of the windows on the mask (e.g., their size, wavelength of transmittance, orientation of polarization) determine the information carried in these optical pulses (e.g., intensity, color, polarization). The structure of a transmitted signal depends on the number and spacing of droplets in the channel. Fourier transformation can deconvolve superimposed signals created by the flow of multiple droplets into the message that a single droplet would transmit. The research described in this contribution explores a new field at the intersection of chemistry, materials science, and information technology: infochemistry.

Quantifying colorimetric assays in paper-based microfluidic devices by measuring the transmission of light through paper.

This article describes a point-of-care (POC) system--comprising a microfluidic, paper-based analytical device (micro-PAD) and a hand-held optical colorimeter--for quantifying the concentration of analytes in biological fluids. The micro-PAD runs colorimetric assays, and consists of paper that has been (i) patterned to expose isolated regions of hydrophilic zones and (ii) wet with an index-matching fluid (e.g., vegetable oil) that is applied using a disposable, plastic sleeve encasement. Measuring transmittance through paper represents a new method of quantitative detection that expands the potential functionality of micro-PADs. This prototype transmittance colorimeter is inexpensive, rugged, and fully self-contained, and thus potentially attractive for use in resource-limited environments and developing countries.

Investigating nanoscale cellular dynamics with cross-sectional spectral domain phase microscopy.

We report on cross-sectional imaging of dynamic biological specimens using a spectral domain phase microscopy (SDPM) system capable of operating at a line rate of 19 kHz. This system combines the time-sensitive capabilities of SDPM with the multi-point acquisition features of related phase-sensitive techniques. The presented phase portraits and B-scan phase images of spontaneously beating embryonic cardiomyocytes and cytoplasmic flow in A. proteus offer insight into the nature and timing of the observed cellular phenomena, demonstrating the utility of this technique for dynamic cell studies.

Spectral domain phase microscopy for local measurements of cytoskeletal rheology in single cells.

We present spectral domain phase microscopy (SDPM) as a new tool for measurements at the cellular scale. SDPM is a functional extension of spectral domain optical coherence tomography that allows for the detection of cellular motions and dynamics with nanometer-scale sensitivity in real time. Our goal was to use SDPM to investigate the mechanical properties of the cytoskeleton of MCF-7 cells. Magnetic tweezers were designed to apply a vertical force to ligand-coated magnetic beads attached to integrin receptors on the cell surfaces. SDPM was used to resolve cell surface motions induced by the applied stresses. The cytoskeletal response to an applied force is shown for both normal cells and those with compromised actin networks due to treatment with Cytochalasin D. The cell response data were fit to several models for cytoskeletal rheology, including one- and two-exponential mechanical models, as well as a power law. Finally, we correlated displacement measurements to physical characteristics of individual cells to better compare properties across many cells, reducing the coefficient of variation of extracted model parameters by up to 50%.

Erratum: Sheathless hydrodynamic positioning of buoyant drops and bubbles inside microchannels [Phys. Rev. E 84, 036302 (2011)].

This corrects the article DOI: 10.1103/PhysRevE.84.036302.

Correction: The magnitude of lift forces acting on drops and bubbles in liquids flowing inside microchannels.

Correction for 'The magnitude of lift forces acting on drops and bubbles in liquids flowing inside microchannels' by Claudiu A. Stan et al., Lab Chip, 2013, 13, 365-376.

Doppler flow imaging of cytoplasmic streaming using spectral domain phase microscopy.

Spectral domain phase microscopy (SDPM) is a function extension of spectral domain optical coherence tomography. SDPM achieves exquisite levels of phase stability by employing common-path interferometry. We discuss the theory and limitations of Doppler flow imaging using SDPM, demonstrate monitoring the thermal contraction of a glass sample with nanometer per second velocity sensitivity, and apply this technique to measurement of cytoplasmic streaming in an Amoeba proteus pseudopod. We observe reversal of cytoplasmic flow induced by extracellular CaCl2, and report results that suggest parabolic flow of cytoplasm in the A. proteus pseudopod.

Label-free and non-contact optical biosensing of glucose with quantum dots.

We present a label-free, optical sensor for biomedical applications based on changes in the visible photoluminescence (PL) of quantum dots in a thin polymer film. Using glucose as the target molecule, the screening of UV excitation due to pre-absorption by the product of an enzymatic assay leads to quenching of the PL of quantum dots (QDs) in a non-contact scheme. The irradiance changes in QD PL indicate quantitatively the level of glucose present. The non-contact nature of the assay prevents surface degradation of the QDs, which yields an efficient, waste-free, cost-effective, portable, and sustainable biosensor with attractive market features. The limit of detection of the demonstrated biosensor is ~3.5 µm, which is competitive with existing contact-based bioassays. In addition, the biosensor operates over the entire clinically relevant range of glucose concentrations of biological fluids including urine and whole blood. The comparable results achieved across a range of cost-affordable detectors, including a spectrophotometer, portable spectrometer, and iPhone camera, suggest that label-free and visible quantification of glucose with QD films can be applied to low-cost, point-of-care biomedical sensing as well as scientific applications in the laboratory for characterizing glucose or other analytes.

Label-free characterization of vitrification-induced morphology changes in single-cell embryos with full-field optical coherence tomography.

Vitrification is an increasingly popular method of embryo cryopreservation that is used in assisted reproductive technology. Although vitrification has high post-thaw survival rates compared to other freezing techniques, its long-term effects on embryo development are still poorly understood. We demonstrate an application of full-field optical coherence tomography (FF-OCT) to visualize the effects of vitrification on live single-cell (2 pronuclear) mouse embryos without harmful labels. Using FF-OCT, we observed that vitrification causes a significant increase in the aggregation of structures within the embryo cytoplasm, consistent with reports in literature based on fluorescence techniques. We quantify the degree of aggregation with an objective metric, the cytoplasmic aggregation (CA) score, and observe a high degree of correlation between the CA scores of FF-OCT images of embryos and of fluorescence images of their mitochondria. Our results indicate that FF-OCT shows promise as a label-free assessment of the effects of vitrification on embryo mitochondria distribution. The CA score provides a quantitative metric to describe the degree to which embryos have been affected by vitrification and could aid clinicians in selecting embryos for transfer.

Fabrication of healthy and disease-mimicking retinal phantoms with tapered foveal pits for optical coherence tomography.

Optical coherence tomography (OCT) has become a standard tool in ophthalmology clinics for diagnosing many retinal diseases. Nonetheless, the technical and clinical communities still lack a standardized phantom that could aid in evaluating and normalizing the many protocols and systems used for diagnosis. Existing retinal phantoms are able to mimic the thickness and scattering properties of the retinal layers but are unable to model the morphology of the foveal pit, particularly the tapering of the retinal layers. This work demonstrates a new fabrication procedure that is capable of reliably and consistently replicating the shape and tapered appearance of the retinal layers near the foveal pit using a combination of spin-coating and replica molding. We characterize the effects of using different mold sizes which enable us to achieve a range of pit dimensions. We also present a modified procedure to replicate two diseased states of the retinal tissue, such as retinal detachment and dry aged-related macular degeneration. The ability to create an anatomically correct foveal pit for healthy and disease-mimicking phantoms will allow for a new standard better suited for intra- and inter-system evaluation and for improved comparison of retinal segmentation algorithms

Using magnetic levitation for non-destructive quality control of plastic parts.

Magnetic levitation (MagLev) enables rapid and non-destructive quality control of plastic parts. The feasibility of MagLev as a method to: i) rapidly assess injection-molded plastic parts for defects during process optimization, ii) monitor the degradation of plastics after exposure to harsh environmental conditions, and iii) detect counterfeit polymers by density is demonstrated.