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Biomedical subjects

Steven L Jacques

Publications and source records attributed to Steven L Jacques.

12 recordsLinked to original sources

Low coherence interferometry of the cochlear partition.

Interferometric measurement of the vibration of the organ of Corti in the isolated guinea pig cochlea was conducted using low-coherence light (1310+/-47 nm wavelength) from a superluminescent diode. The short coherence length of the light source localized measurements along the axial direction to within a approximately 10-microm window (in tissue), even when using a low numerical-aperture lens. The ability to accomplish this is important because measurement of the vibration of the basal-turn organ of Corti is generally done via a small hole in the bone of the cochlea, which effectively limits the numerical aperture. The axial localization, combined with the inherent sensitivity of the method, allowed distinct measurements of the basilar membrane (BM) and the putative reticular lamina (RL) vibration using only the native tissue reflectance, that is without requiring the use of reflective particles. The system was first operated in a scanning mode as an optical coherence tomography (OCT) system to yield an image of the organ of Corti. The reflectance of intensity from the BM and RL was 8x10(-5) and 8x10(-6), respectively. The internal structure between the BM and RL presented a variable reflectivity of about 10(-7). A mirror would define a reflectance of 1.00. Then the instrument was operated as a homodyne interferometer to measure the displacement of either the BM or RL. Vibration at 16 kHz was induced by a piezoelectric actuator, causing whole movement of a dissected cochlea. After calibration of the system, we demonstrated clear measurement of mechanically driven vibration for both the BM and RL of 0.30 nm above a noise floor equivalent to 0.03 nm. OCT interferometry, when adapted for in vivo organ of Corti measurements, appears suitable to determine the micromechanical vibration of cells and tissue elements of the organ.

Animals↗

Simultaneous compensation for spatial and temporal dispersion of acousto-optical deflectors for two-dimensional scanning with a single prism.

The dispersive nature of the acousto-optical deflector (AOD) presents a challenge to applications of two sequential orthogonal AODs (a two-dimensional AOD) as XY scanners in multiphoton microscopy. Introducing a prism before the two-dimensional (2D) AOD allows both temporal and spatial dispersion to be compensated for simultaneously. A 90 fs laser pulse was broadened to 572 fs without compensation, and 143 fs with compensation. The ratio of long axis to short axis of the exiting laser beam spot was 3.50 without compensation and 1.14 with compensation. The insertion loss was 37%. Two-photon fluorescence microscopy used the compensated 2D AOD scanner to image a fluorescent microsphere, which improves signal intensity -15-fold compared with the uncompensated scanner.

Artifacts↗

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.

Algorithms↗

Relationship between damaged fraction and reflected spectra of denaturing tissues.

BACKGROUND AND OBJECTIVES: During thermal therapy of tissue, such as induced by microwave heating, the initiation of denaturation should be monitored for proper thermal dosage. Additionally, denaturation should be confined to the pathologic volume, while preserving surrounding healthy tissue. The relationship between the damaged fraction and reflected spectra of denaturing tissues was investigated for a variation of the temperature of the tissues. STUDY DESIGN/MATERIALS AND METHODS: Denaturation of muscle, liver, and milk was studied in vitro by measuring the temperature-varying reflectance spectrum as heating occurs. A high-resolution fiber optic spectrometer was used to measure the reflectance changes. Temperature was monitored using a thermocouple embedded within the tissue along the side of the optical fiber probe. RESULTS: The values of average free energy to initiate denaturation in muscle and liver at about 60 degrees C were 94.8 and 96.3 kJ/mole, respectively. The reflectance spectra increased in amplitude for muscle and liver, and the peak shifted from approximately 700 to 720 nm in accordance with the damage fraction of tissue. The reflectance spectrum for milk was essentially unchanged. CONCLUSIONS: Spectral changes from heated muscle and liver reflect denaturation of proteins contained therein. The spectral information at 800 nm can be used to determine the average free energy for the initiation of denaturation.

Animals↗

Imaging melanoma in a murine model using reflectance-mode confocal scanning laser microscopy and polarized light imaging.

The light-scattering properties of cutaneous tissues provide optical contrast for imaging the presence and depth of pigmented melanoma in a highly pigmented murine model, the C57/B6 mouse. Early lesions are difficult to identify when viewing black lesions on a black mouse. Two methods were used to image early lesions in this model. (1) A reflectance-mode confocal scanning laser microscope (rCSLM) was built to provide horizontal images (x-y at depth z) and transverse images (x-z at position y) non-invasively in the living mouse. (2) A polarized light imaging (PLI) camera was built using a linearly polarized white light source that viewed the skin through an analyzing linear polarizer oriented either parallel or perpendicular to the illumination's polarization to yield two images, "PAR" and "PER," respectively. The difference image, PAR-PER, eliminated multiply scattered light and yielded an image of the superficial but subsurface tissues based only on photons scattered once or a few times so as to retain their polarization. rCSLM could image melanoma lesions developing below the epidermis. PLI could distinguish superficial from deeper melanoma lesions because the melanin of the superficial lesions attenuated the PAR-PER image, whereas deeper lesions failed to attenuate the PAR-PER image.

Animals↗

Collection efficiency of a single optical fiber in turbid media.

If a single optical fiber is used for both delivery and collection of light, two major factors affect the measurement of collected light: (1) the light transport in the medium that describes the amount of light that returns to the fiber and (2) the light coupling to the optical fiber that depends on the angular distribution of photons entering the fiber. We focus on the importance of the latter factor and describe how the efficiency of the coupling depends on the optical properties of the medium. For highly scattering tissues, the efficiency is well predicted by the numerical aperture (NA) of the fiber. For lower scattering, such as in soft tissues, photons arrive at the fiber from deeper depths, and the coupling efficiency could increase twofold to threefold above that predicted by the NA.

Fiber Optic Technology↗

Clinical testing of a photoacoustic probe for port wine stain depth determination.

BACKGROUND AND OBJECTIVE: Successful laser treatment of port wine stain (PWS) birthmarks requires knowledge of lesion geometry. Laser parameters, such as pulse duration, wavelength, and radiant exposure, and other treatment parameters, such as cryogen spurt duration, need to be optimized according to epidermal melanin content and lesion depth. We designed, constructed, and clinically tested a photoacoustic probe for PWS depth determination. STUDY DESIGN/MATERIALS AND METHODS: Energy from a frequency-doubled, Nd:YAG laser (lambda=532 nm, tau(p)=4 nanoseconds) was coupled into two 1,500 mum optical fibers fitted into an acrylic handpiece containing a piezoelectric acoustic detector. Laser light induced photoacoustic waves in tissue phantoms and a patient's PWS. The photoacoustic propagation time was used to calculate the depth of the embedded absorbers and PWS lesion. RESULTS: Calculated chromophore depths in tissue phantoms were within 10% of the actual depths of the phantoms. PWS depths were calculated as the sum of the epidermal thickness, determined by optical coherence tomography (OCT), and the epidermal-to-PWS thickness, determined photoacoustically. PWS depths were all in the range of 310-570 microm. The experimentally determined PWS depths were within 20% of those measured by optical Doppler tomography (ODT). CONCLUSIONS: PWS lesion depth can be determined by a photoacoustic method that utilizes acoustic propagation time.

Dermatology↗

Imaging skin pathology with polarized light.

Linearly polarized light that illuminates skin is backscattered by superficial layers and rapidly depolarized by birefringent collagen fibers. It is possible to distinguish such superficially backscattered light from the total diffusely reflected light that is dominated by light penetrating deeply into the dermis. The method involves acquisition of two images through an analyzing linear polarizer in front of the camera, one image (I(par)) acquired with the analyzer oriented parallel to the polarization of illumination and one image (I(per)) acquired with the analyzer oriented perpendicular to the illumination. An image based on the polarization ratio, Pol=(I(par)-I(per))/(I(par)+I(per)), is created. This paper compares normal light images, represented by I(per), and Pol images of various skin pathologies in a pilot clinical study using incoherent visible-spectrum light. Images include pigmented skin sites (freckle, tattoo, pigmented nevi) and unpigmented skin sites [nonpigmented intradermal nevus, neurofibroma, actinic keratosis, malignant basal cell carcinoma, squamous cell carcinoma, vascular abnormality (venous lake), burn scar]. Images of a shadow cast from a razor blade onto a forearm skin site illustrate the behavior of Pol values near the shadow edge. Near the shadow edge, Pol approximately doubles in value because no I(per) photons are superficially scattered into the shadow-edge pixels by the shadow region while I(par) photons are directly backscattered from the superficial layer of these pixels. This result suggests that the point spread function in skin for cross-talk between Pol pixels has a half-width-half-max of about 390 microm.

Adult↗

Confocal fluorescence spectroscopy of subcutaneous cartilage expressing green fluorescent protein versus cutaneous collagen autofluorescence.

Optically monitoring the expression of green fluorescent protein (GFP) in the cartilage underlying the skin of a mouse allows tracking the expression of the chondrocyte phenotype. This paper considers how confocal microscopy with spectral detection can sense GFP fluorescence in the cartilage despite light scattering and collagen autofluorescence from the overlying skin. An in vivo experiment tested the abilities of a topical optical fiber measurement and a confocal microscope measurement to detect GFP in cartilage under the skin versus the collagen autofluorescence. An ex vivo experiment tested the ability of a confocal microscope without and with its pinhole to detect a fluorescent microsphere underneath an ex vivo skin layer versus the collagen autofluorescence. In both systems, spectroscopic detection followed by linear analysis allowed spectral discrimination of collagen autofluorescence (M(C)) and the subdermal green fluorescence (M(G)) due to either GFP or the microsphere. Contrast was defined as M(G)/(M(G)+M(C)). The in vivo contrast for GFP using optical fiber and confocal measurements was 0.16 and 0.92, respectively. The ex vivo contrast for a fluorescent microsphere using a confocal system without and with a pinhole was 0.13 and 0.48, respectively. The study demonstrates that a topical optical fiber measurement is affected by collagen autofluorescence, while a confocal microscope can detect subdermal fluorescence while rejecting collagen autofluorescence.

Animals↗

Design, testing, and clinical studies of a handheld polarized light camera.

Polarized light imaging has been used to detect the borders of skin cancer and facilitate assessment of cancer boundaries. A design for an inexpensive handheld polarized camera is presented and clinical images acquired with this prototype are shown. The camera is built with two universal serial bus (USB) color video cameras, a polarizing beamsplitter cube, and a 4x objective lens. Illumination is provided by three white LEDs and a sheet polarizer. Horizontal and vertical linearly polarized reflected images are processed at 7 frames/s and a resulting polarized image is displayed on screen. We compare the performances of cheap USB camera and a 16-bit electronically cooled camera. Dark noise and image repeatability are compared. In both cases, the 16-bit camera outperforms the USB cameras. Despite these limitations, the results obtained with this USB prototype are very satisfactory. Examples of polarized images of lesions taken prior to surgery are presented.

Computer Communication Networks↗

In vivo optical reflectance imaging of spreading depression waves in rat brain with and without focal cerebral ischemia.

Spreading depression (SD) waves occur in focal cerebral ischemia of the brain. Optical reflectance imaging at 550 +/- 10-nm wavelength using a charge-coupled device (CCD) camera, called optical intrinsic signal imaging (OISI) in the neuroscience community, provides high resolution imaging of SD waves based on changes in blood perfusion. We present optical images of SD waves in normal rat brain induced by a pinprick, and the spontaneous SD waves that follow middle cerebral artery occlusion (MCAO). The images of change in reflectance are calculated as A = (I-I(o))I(o), where I is pixel intensity as some timepoint and I(o) is the initial intensity just prior to an SD wave. Difference images B = [I(i)-I(i-1)]I(o), where I(i) is the image at time i and I(i-1) is the previous image at time i-1 (a 6.4-s interval), significantly sharpen the boundaries between leading and trailing edges of the SD wave. Maximum rate-of-change images C = max(B) display the maximum pixel value of B within the duration of a single SD wave, and provide an image that visualizes the entire penumbra. The penumbra appear bright due to a rapid drop in perfusion, while the normal brain and infarct area appear dark.

Animals↗

Ratio of entropy to enthalpy in thermal transitions in biological tissues.

Thermal transitions in biological tissues that have been reported in the literature are summarized in terms of the apparent molar entropy (DeltaS) and molar enthalpy (DeltaH) involved in the transition. A plot of DeltaS versus DeltaH for all the data yields a straight line, consistent with the definition of free energy, DeltaG=DeltaH+TDeltaS. Various bonds may be involved in cooperative bond breakage during thermal transitions; however, for the sake of description, the equivalent number of cooperative hydrogen bonds can be cited. Most of the tissue data behave as if 10 to 20 hydrogen bonds are cooperatively broken during coagulation, with one transition, the expression of heat shock protein, involving 90 cooperative hydrogen bonds. The data are consistent with DeltaS=a+bDeltaH, where a=-327.5 J(mol K) and b=31.47 x 10(-4) K(-1). If each additional hydrogen bond adds 19 x 10(3) Jmol to DeltaH, then each additional bond adds 59.8 J(mol K) to DeltaS. Hence, the dynamics of irreversible thermal transitions can be described in terms of one free parameter, the apparent number of cooperative hydrogen bonds broken during the transition.

Animals↗