Timothy R. Hillman

In vivo dynamic optical coherence elastography using a ring actuator

We present a novel sample arm arrangement for dynamic optical coherence elastography based on excitation by a ring actuator. The actuator enables coincident excitation and imaging to be performed on a sample, facilitating in vivo operation. Sub-micrometer vibrations in the audio frequency range were coupled to samples that were imaged using optical coherence tomography. The resulting vibration amplitude and microstrain maps are presented for bilayer silicone phantoms and multiple skin sites on a human subject. Contrast based on the differing elastic properties is shown, notably between the epidermis and dermis. The results constitute the first demonstration of a practical means of performing in vivo dynamic optical coherence elastography on a human subject.

Optical measurement of biomechanical properties of individual erythrocytes from a sickle cell patient.

Sickle cell disease (SCD) is characterized by the abnormal deformation of red blood cells (RBCs) in the deoxygenated condition, as their elongated shape leads to compromised circulation. The pathophysiology of SCD is influenced by both the biomechanical properties of RBCs and their hemodynamic properties in the microvasculature. A major challenge in the study of SCD involves accurate characterization of the biomechanical properties of individual RBCs with minimum sample perturbation. Here we report the biomechanical properties of individual RBCs from a SCD patient using a non-invasive laser interferometric technique. We optically measure the dynamic membrane fluctuations of RBCs. The measurements are analyzed with a previously validated membrane model to retrieve key mechanical properties of the cells: bending modulus; shear modulus; area expansion modulus; and cytoplasmic viscosity. We find that high cytoplasmic viscosity at ambient oxygen concentration is principally responsible for the significantly decreased dynamic membrane fluctuations in RBCs with SCD, and that the mechanical properties of the membrane cortex of irreversibly sickled cells (ISCs) are different from those of the other types of RBCs in SCD.

Digital optical phase conjugation for delivering two-dimensional images through turbid media

Optical transmission through complex media such as biological tissue is fundamentally limited by multiple light scattering. Precise control of the optical wavefield potentially holds the key to advancing a broad range of light-based techniques and applications for imaging or optical delivery. We present a simple and robust digital optical phase conjugation (DOPC) implementation for suppressing multiple light scattering. Utilizing wavefront shaping via a spatial light modulator (SLM), we demonstrate its turbidity-suppression capability by reconstructing the image of a complex two-dimensional wide-field target through a highly scattering medium. Employing an interferometer with a Sagnac-like ring design, we successfully overcome the challenging alignment and wavefront-matching constraints in DOPC, reflecting the requirement that the forward- and reverse-propagation paths through the turbid medium be identical. By measuring the output response to digital distortion of the SLM write pattern, we validate the sub-wavelength sensitivity of the system.

Correlation of static speckle with sample properties in optical coherence tomography

We present theoretical calculations, based on a random phasor sum model, which show that the optical coherence tomography speckle contrast ratio is dependent on the local density of scattering particles in a sample, provided that the effective number of scatterers in the probed volume is less than about five. We confirm these theoretical predictions experimentally, using suspensions of microspheres in water. The observed contrast ratios vary in value from the Rayleigh limit of 0.52 to in excess of 2, suggesting that the contrast ratio could be useful in optical coherence tomography, particularly when imaging in ultrahigh-resolution regimes.

The effect of water dispersion and absorption on axial resolution in ultrahigh-resolution optical coherence tomography

We examine the effects of dispersion and absorption in ultrahigh-resolution optical coherence tomography (OCT), particularly the necessity to compensate for high dispersion orders in order to narrow the axial point-spread function envelope. We present a numerical expansion in which the impact of the various dispersion orders is quantified; absorption effects are evaluated numerically. Assuming a Gaussian source spectrum (in the optical frequency domain), we focus on imaging through water as a first approximation to biological materials. Both dispersion and absorption are found to be most significant for wavelengths above ~ 1microm, so that optimizing the system effective resolution (ER) requires choosing an operating wavelength below this limit. As an example, for 1-microm source resolution (FWHM), and propagation through a 1-mm water cell, if up to third-order dispersion compensation is applied, then the optimal center wavelength is 0.8microm, which generates an ER of 1.5microm (in air). The incorporation of additional bandwidth yields no ER improvement, due to uncompensated high-order dispersion and long-wavelength absorption.

Speckle reduction in optical coherence tomography by strain compounding

We present a speckle reduction technique for optical coherence tomography based on strain compounding. Decorrelation is introduced between B-scans by altering the samples strain. A theoretical description of the technique, based on a transfer-function formalism, and experimental results on silicone phantoms are presented. Nearly complete decorrelation between successive B-scan speckle patterns was observed for a variation in strain of 0.045. Strain compounding by averaging nine B-scans, with 0.003 strain increments between them, resulted in a 1.5-fold reduction in speckle contrast ratio.

Synthetic Fourier transform light scattering.

We present synthetic Fourier transform light scattering, a method for measuring extended angle-resolved light scattering (ARLS) from individual microscopic samples. By measuring the light fields scattered from the sample plane and numerically synthesizing them in Fourier space, the angle range of the ARLS patterns is extended up to twice the numerical aperture of the imaging system with unprecedented sensitivity and precision. Extended ARLS patterns of individual microscopic polystyrene beads, healthy human red blood cells (RBCs), and Plasmodium falciparum-parasitized RBCs are presented.

Coherent aperture-synthesis, wide-field, high-resolution holographic microscopy of biological tissue

We show for the first time, to our knowledge, high-resolution wide-field images of biological samples recorded using coherent aperture-synthesis Fourier holography. To achieve this, we combined off-axis plane-wave polarized illumination with an axial sample rotation and polarization-sensitive collection of backscattered light. We synthesized 180 Fourier holograms using an efficient postdetection phase-matching correlation scheme. The result was an annular spatial frequency-space synthetic aperture (NA=0.93) with an effective area 25 times larger than that due to a single hologram. A high-resolution high-contrast microscopic reconstruction of biological tissue was computed over a sample area of 9 mm(2) from holograms acquired at 34 mm working distance.

Spatially resolved Fourier holographic light scattering angular spectroscopy

We utilize Fourier-holographic light scattering angular spectroscopy to record the spatially resolved complex angular scattering spectra of samples over wide fields of view in a single or few image captures. Without resolving individual scatterers, we are able to generate spatially-resolved particle size maps for samples composed of spherical scatterers, by comparing generated spectra with Mie-theory predictions. We present a theoretical discussion of the fundamental principles of our technique and, in addition to the sphere samples, apply it experimentally to a biological sample which comprises red blood cells. Our method could possibly represent an efficient alternative to the time-consuming and laborious conventional procedure in light microscopy of image tiling and inspection, for the characterization of microscopic morphology over wide fields of view.

Single-shot quantitative dispersion phase microscopy

We present an imaging modality capable of providing high-speed optical dispersion measurements of live cells. The technique permits wide-field measurement of the optical phase shifts introduced by a sample for multiple discrete wavelengths in a single image capture. Utilizing spatial modulation and the wavelength dependence of the interference-fringe spacing, average refractive index as a function of wavelength is obtained, yielding optical dispersion measurements of the sample under observation. Because of its simple and low-cost design, the technique can be readily integrated into a standard microscope to collect additional diagnostic information about biological cells.