Widefield spatial frequency domain imaging of tissue oxygen saturation (StO2), superficial (HbT1) and deep (HbT2) hemoglobin concentrations was performed on an arterial-venous insufficiency created by a forearm vascular occlusion test in 15 healthy subjects. Measured StO2 and HbT2 were significantly higher for regions of interest having underlying vessels compared to those without (microvasclature) at each time point (p < 0.01). During occlusion, the decrease in StO2 was greater for microvascular regions. These spatially resolved measurements suggest that image-based analysis is important as probe measurements can be confounded by probe location on the heterogeneous tissue.
Spatial frequency domain imaging (SFDI) has witnessed very rapid growth over the last decade, owing to its unique capabilities for imaging optical properties and chromophores over a large field-of-view and in a rapid manner. We provide a comprehensive review of the principles of this imaging method as of 2019, review the modeling of light propagation in this domain, describe acquisition methods, provide an understanding of the various implementations and their practical limitations, and finally review applications that have been published in the literature. Importantly, we also introduce a group effort by several key actors in the field for the dissemination of SFDI, including publications, advice in hardware and implementations, and processing code, all freely available online.
Imaging Cherenkov emission during radiotherapy permits real-time visualization of external beam delivery on superficial tissue. This signal is linear with absorbed dose in homogeneous media, indicating potential for quantitative dosimetry. In humans, the inherent heterogeneity of tissue optical properties (primarily from blood and skin pigment) distorts the linearity between detected Cherenkov signal and absorbed dose. We examine the potential to correct for superficial vasculature using spatial frequency domain imaging (SFDI) to map tissue optical properties for large fields of view. In phantoms, applying intensity corrections to simulate blood vessels improves Cherenkov image (CI) negative contrast by 24% for a vessel 1.9-mm-in diameter. In human trials, SFDI and CI are acquired for women undergoing whole breast radiotherapy. Applied corrections reduce heterogeneity due to vasculature within the sampling limits of the SFDI from a 22% difference as compared to the treatment plan, down to 6% in one region and from 14% down to 4% in another region. The optimal use for this combined imaging system approach is to correct for small heterogeneities such as superficial blood vessels or for interpatient variations in blood/melanin content such that the corrected CI more closely represents the surface dose delivered.
We introduce a tomographic approach for three-dimensional imaging of evoked hemodynamic activity, using broadband illumination and diffuse optical tomography (DOT) image reconstruction. Changes in diffuse reflectance in the rat somatosensory cortex due to stimulation of a single whisker were imaged at a frame rate of 5 Hz using a hyperspectral image mapping spectrometer. In each frame, images in 38 wavelength bands from 484 to 652 nm were acquired simultaneously. For data analysis, we developed a hyperspectral DOT algorithm that used the Rytov approximation to quantify changes in tissue concentration of oxyhemoglobin (ctHbO2) and deoxyhemoglobin (ctHb) in three dimensions. Using this algorithm, the maximum changes in ctHbO2 and ctHb were found to occur at 0.29±0.02 and 0.66±0.04 mm beneath the surface of the cortex, respectively. Rytov tomographic reconstructions revealed maximal spatially localized increases and decreases in ctHbO2 and ctHb of 321±53 and 555±96 nM, respectively, with these maximum changes occurring at 4±0.2 s poststimulus. The localized optical signals from the Rytov approximation were greater than those from modified Beer–Lambert, likely due in part to the inability of planar reflectance to account for partial volume effects.
The standard of care for clinical assessment of burn severity and extent lacks a quantitative measurement. In this work, spatial frequency domain imaging (SFDI) was used to measure 48 thermal burns of graded severity (superficial partial, deep partial, and full thickness) in a porcine model. Functional (total hemoglobin and tissue oxygen saturation) and structural parameters (tissue scattering) derived from the SFDI measurements were monitored over 72 h for each burn type and compared to gold standard histological measurements of burn depth. Tissue oxygen saturation (stO2) and total hemoglobin (ctHbT) differentiated superficial partial thickness burns from more severe burn types after 2 and 72 h, respectively (p<0.01), but were unable to differentiate deep partial from full thickness wounds in the first 72 h. Tissue scattering parameters separated superficial burns from all burn types immediately after injury (p<0.01), and separated all three burn types from each other after 24 h (p<0.01). Tissue scattering parameters also showed a strong negative correlation to histological burn depth as measured by vimentin immunostain (r2>0.89). These results show promise for the use of SFDI-derived tissue scattering as a correlation to burn depth and the potential to assess burn depth via a combination of SFDI functional and structural parameters.
Recently, digital photography has become an efficient and economic method to assist dermatologists in monitoring skin characteristics. Although this technology has advanced a great deal in resolution and costs, conventional digital cameras continue to only provide qualitative recording of color information. To address this issue, we are developing a compact, quantitative skin imaging camera by employing spatial frequency domain imaging (SFDI), a non-contact approach for determining tissue optical properties over a wide field-of-view. SFDI uses knowledge of optical properties at multiple wavelengths to recover concentrations of tissue constituents such as oxy/deoxy-hemoglobin, water, and melanin. This method has been well researched and presented in laboratory and research settings. The next step in the development of SFDI systems is to make typical systems compact and cheaper using commercial components. We present our findings by performing a component-by-component analysis of key SFDI system components including light sources, projectors, and cameras.
Spatial Frequency Domain Imaging (SFDI) is a non-contact imaging method that uses multiple frequency spatial
illumination to generate two dimensional maps of tissue optical properties (absorption and reduced scattering) and
chromophore concentrations. We present phantom validation and pilot clinical data of a deployed light-emitting diode
(LED) based system. The system employs four LED wavelengths (658 nm, 730 nm, 850 nm, 970 nm) to quantitatively
assess tissue health by measurement of common tissue constituents. Phantom validation results and maps of oxyhemoglobin,
deoxy-hemoglobin, water content, reduced scattering, and surface topography will be presented for pilot
studies assessing burn severity and efficacy of port wine stain treatment.
Oxygenation measurements are widely used in patient care. However, most clinically available instruments currently consist of contact probes that only provide global monitoring of the patient (e.g., pulse oximetry probes) or local monitoring of small areas (e.g., spectroscopy-based probes). Visualization of oxygenation over large areas of tissue, without a priori knowledge of the location of defects, has the potential to improve patient management in many surgical and critical care applications. In this study, we present a clinically compatible multispectral spatial frequency domain imaging (SFDI) system optimized for surgical oxygenation imaging. This system was used to image tissue oxygenation over a large area (16×12 cm) and was validated during preclinical studies by comparing results obtained with an FDA-approved clinical oxygenation probe. Skin flap, bowel, and liver vascular occlusion experiments were performed on Yorkshire pigs and demonstrated that over the course of the experiment, relative changes in oxygen saturation measured using SFDI had an accuracy within 10% of those made using the FDA-approved device. Finally, the new SFDI system was translated to the clinic in a first-in-human pilot study that imaged skin flap oxygenation during reconstructive breast surgery. Overall, this study lays the foundation for clinical translation of endogenous contrast imaging using SFDI.
Introduction: Two major disadvantages of currently available oxygenation probes are the need for contact with the skin
and long measurement stabilization times. A novel oxygenation imaging device based on spatial frequency domain and
spectral principles has been designed, validated preclinically on pigs, and validated clinically on humans. Importantly,
this imaging system has been designed to operate under the rigorous conditions of an operating room. Materials and
Methods: Optical properties reconstruction and wavelength selection have been optimized to allow fast and reliable
oxyhemoglobin and deoxyhemoglobin imaging under realistic conditions. In vivo preclinical validation against
commercially available contact oxygenation probes was performed on pigs undergoing arterial and venous occlusions.
Finally, the device was used clinically to image skin flap oxygenation during a pilot study on women undergoing breast
reconstruction after mastectomy. Results: A novel illumination head containing a spatial light modulator (SLM) and a
novel fiber-coupled high power light source were constructed. Preclinical experiments showed similar values between
local probes and the oxygenation imaging system, with measurement times of the new system being < 500 msec. During
pilot clinical studies, the imaging system was able to provide near real-time oxyHb, deoxyHb, and saturation
measurements over large fields of view (> 300 cm2). Conclusion: A novel optical-based oxygenation imaging system has
the potential to replace contact probes during human surgery and to provide quantitative, wide-field measurements in
near real-time.
We present a rapid, noncontact imaging technique which can obtain the spectrally- and spatially-resolved scattering
and absorption coefficients of a turbid medium. The measurement involves combining a spatially modulated
illumination pattern with a snapshot imaging spectrometer for measurement. After capture of an (x, y, λ)
datacube, an image demodulation scheme is applied in post-processing to obtain the spatial maps of diffuse
reflectance, absorption coefficient, and reduced scattering coefficient. The resulting system is used to dynamic
maps (in 1 s intervals) of the brain's intrinsic optical signal.
Spatial frequency-domain imaging (SFDI) utilizes multiple-frequency structured illumination and model-based computation to generate two-dimensional maps of tissue absorption and scattering properties. SFDI absorption data are measured at multiple wavelengths and used to fit for the tissue concentration of intrinsic chromophores in each pixel. This is done with a priori knowledge of the basis spectra of common tissue chromophores, such as oxyhemoglobin (ctO2Hb), deoxyhemoglobin (ctHHb), water (ctH2O), and bulk lipid. The quality of in vivo SFDI fits for the hemoglobin parameters ctO2Hb and ctHHb is dependent on wavelength selection, fitting parameters, and acquisition rate. The latter is critical because SFDI acquisition time is up to six times longer than planar two-wavelength multispectral imaging due to projection of multiple-frequency spatial patterns. Thus, motion artifact during in vivo measurements compromises the quality of the reconstruction. Optimal wavelength selection is examined through matrix decomposition of basis spectra, simulation of data, and dynamic in vivo measurements of a human forearm during cuff occlusion. Fitting parameters that minimize cross-talk from additional tissue chromophores, such as water and lipid, are determined. On the basis of this work, a wavelength pair of 670 nm/850 nm is determined to be the optimal two-wavelength combination for in vivo hemodynamic tissue measurements provided that assumptions for water and lipid fractions are made in the fitting process. In our SFDI case study, wavelength optimization reduces acquisition time over 30-fold to 1.5s compared to 50s for a full 34-wavelength acquisition. The wavelength optimization enables dynamic imaging of arterial occlusions with improved spatial resolution due to reduction of motion artifacts.
Sinusoidally structured illumination is used in concert with a phantom-based lookup-table (LUT) to map wide-field optical properties in turbid media with reduced albedos as low as 0.44. A key advantage of the lookup-table approach is the ability to measure the absorption (µa) and reduced scattering coefficients (µ′s) over a much broader range of values than permitted by current diffusion theory methods. Through calibration with a single reflectance standard, the LUT can extract µ′s from 0.8 to 2.4 mm-1 with an average root-mean-square (rms) error of 7% and extract µa from 0 to 1.0 mm-1 with an average rms error of 6%. The LUT is based solely on measurements of two parameters, reflectance R and modulation M at an illumination period of 10 mm. A single set of three phase-shifted images is sufficient to measure both M and R, which are then used to generate maps of absorption and scattering by referencing the LUT. We establish empirically that each pair (M,R) maps uniquely to only one pair of (µ′s,µa) and report that the phase function (i.e., size) of the scatterers can influence the accuracy of optical property extraction.
We introduce a noncontact imaging method utilizing multifrequency structured illumination for improving lateral and axial resolution and contrast of fluorescent molecular probes in thick, multiple-scattering tissue phantoms. The method can be implemented rapidly using a spatial light modulator and a simple image demodulation scheme similar to structured light microscopy in the diffraction regime. However, imaging is performed in the multiple-scattering regime utilizing spatially modulated scalar photon density waves. We demonstrate that by increasing the structured light spatial frequency, fluorescence from deeper structures is suppressed and signals from more superficial objects enhanced. By measuring the spatial frequency dependence of fluorescence, background can be reduced by localizing the signal to a buried fluorescent object. Overall, signal-to-background ratio (SBR) and resolution improvements are dependent on spatial frequency and object depth/dimension with as much as sevenfold improvement in SBR and 33% improvement in resolution for ~1-mm objects buried 3 mm below the surface in tissue-like media with fluorescent background.
We describe a noncontact profile correction technique for quantitative, wide-field optical measurement of tissue absorption (µa) and reduced scattering (µ) coefficients, based on geometric correction of the sample's Lambertian (diffuse) reflectance intensity. Because the projection of structured light onto an object is the basis for both phase-shifting profilometry and modulated imaging, we were able to develop a single instrument capable of performing both techniques. In so doing, the surface of the three-dimensional object could be acquired and used to extract the object's optical properties. The optical properties of flat polydimethylsiloxane (silicone) phantoms with homogenous tissue-like optical properties were extracted, with and without profilometry correction, after vertical translation and tilting of the phantoms at various angles. Objects having a complex shape, including a hemispheric silicone phantom and human fingers, were acquired and similarly processed, with vascular constriction of a finger being readily detectable through changes in its optical properties. Using profilometry correction, the accuracy of extracted absorption and reduced scattering coefficients improved from two- to ten-fold for surfaces having height variations as much as 3 cm and tilt angles as high as 40 deg. These data lay the foundation for employing structured light for quantitative imaging during surgery.
The cochlea is the mammalian organ of hearing. Its predominant vibratory element, the basilar membrane, is tonotopically tuned, based on the spatial variation of its mass and stiffness. The constituent collagen fibers of the basilar membrane affect its stiffness. Laser irradiation can induce collagen remodeling and deposition in various tissues. We tested whether similar effects could be induced within the basilar membrane. Trypan blue was perfused into the scala tympani of anesthetized mice to stain the basilar membrane. We then irradiated the cochleas with a 694-nm pulsed ruby laser at 15 or 180 J/cm2. The mice were sacrificed 14 to 16 days later and collagen organization was studied. Polarization microscopy revealed that laser irradiation increased the birefringence within the basilar membrane in a dose-dependent manner. Electron microscopy demonstrated an increase in the density of collagen fibers and the deposition of new fibrils between collagen fibers after laser irradiation. As an assessment of hearing, auditory brainstem response (ABR) thresholds were found to increase moderately after 15 J/cm2 and substantially after 180 J/cm2. Our results demonstrate that collagen remodeling and new collagen deposition occurs within the basilar membrane after laser irradiation in a similar fashion to that found in other tissues.
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