In reconstructive surgery, the ability to detect blood flow interruptions to grafted tissue represents a critical step in preventing postsurgical complications. We have developed and pilot tested a compact, fiber-based device that combines two complimentary modalities—diffuse correlation spectroscopy (DCS) and diffuse reflectance spectroscopy—to quantitatively monitor blood perfusion. We present a proof-of-concept study on an in vivo porcine model (n=8). With a controllable arterial blood flow supply, occlusion studies (n=4) were performed on surgically isolated free flaps while the device simultaneously monitored blood flow through the supplying artery as well as flap perfusion from three orientations: the distal side of the flap and two transdermal channels. Further studies featuring long-term monitoring, arterial failure simulations, and venous failure simulations were performed on flaps that had undergone an anastomosis procedure (n=4). Additionally, benchtop verification of the DCS system was performed on liquid flow phantoms. Data revealed relationships between diffuse optical measures and state of occlusion as well as the ability to detect arterial and venous compromise. The compact construction of the device, along with its noninvasive and quantitative nature, would make this technology suitable for clinical translation.
Optical tissue phantoms are necessary for instrument benchmarking and providing a consistent baseline for experiments in various fields of tissue spectroscopy, including diffuse correlation spectroscopy (DCS). To provide the most useful comparisons, a phantom would ideally mimic tissue as closely as possible, including the geometry of static and dynamic scatterers. A branching design that keeps the capillary cross section constant ensures that the same flow velocity is found throughout the phantom while allowing for single input and output fittings to feed all of the capillaries simultaneously. The direction of each capillary is randomized every few millimeters by randomly allocating 2 by 2 "twisting" squares within each layer. These squares swap the locations of four adjacent artificial capillaries either clockwise or counterclockwise. Numerical simulations were used to verify the random walk-like behavior of the capillary paths resulting from this pattern. This is a step toward replicating the randomly varying directionality of actual capillaries. This design was verified by taking DCS measurements at different flow rates of Intralipid through the phantom, demonstrating the effect of the flow rate on the characteristic decay time of the autocorrelation.
It is essential to monitor tissue perfusion during and after reconstructive surgery, as restricted blood flow can result in graft failures. Current clinical procedures are insufficient to monitor tissue perfusion, as they are intermittent and often subjective. To address this unmet clinical need, a compact, low-cost, multimodal diffuse correlation spectroscopy and diffuse reflectance spectroscopy system was developed. We verified system performance via tissue phantoms and experimental protocols for rigorous bench testing. Quantitative data analysis methods were employed and tested to enable the extraction of tissue perfusion parameters. This design verification study assures data integrity in future in vivo studies.
In reconstructive surgery, impeded blood flow in microvascular free flaps due to a compromise in arterial or venous patency secondary to blood clots or vessel spasms can rapidly result in flap failures. Thus, the ability to detect changes in microvascular free flaps is critical. In this paper, we report progress on in vivo pre-clinical testing of a compact, multimodal, fiber-based diffuse correlation and reflectance spectroscopy system designed to quantitatively monitor tissue perfusion in a porcine model’s surgically-grafted free flap. We also describe the device’s sensitivity to incremental blood flow changes and discuss the prospects for continuous perfusion monitoring in future clinical translational studies.
In reconstructive surgery, tissue perfusion/vessel patency is critical to the success of microvascular free tissue flaps. Early detection of flap failure secondary to compromise of vascular perfusion would significantly increase the chances of flap salvage. We have developed a compact, clinically-compatible monitoring system to enable automated, minimally-invasive, continuous, and quantitative assessment of flap viability/perfusion. We tested the system’s continuous monitoring capability during extended non-recovery surgery using an in vivo porcine free flap model. Initial results indicated that the system could assess flap viability/perfusion in a quantitative and continuous manner. With proven performance, the compact form constructed with cost-effective components would make this system suitable for clinical translation.
Currently the diagnosis of hemorrhagic shock is essentially clinical, relying on the expertise of nurses and doctors. One of the first measurable physiological changes that marks the onset of hemorrhagic shock is a decrease in capillary blood flow. Diffuse correlation spectroscopy (DCS) quantifies this decrease. DCS collects and analyzes multiply scattered, coherent, near infrared light to assess relative blood flow. This work presents a preliminary study using a DCS instrument with human subjects undergoing a lower body negative pressure (LBNP) protocol. This work builds on previous successful DCS instrumentation development and we believe it represents progress toward understanding how DCS can be used in a clinical setting.
Diffuse correlation spectroscopy (DCS) is a technique which enables powerful and robust non-invasive optical studies of tissue micro-circulation and vascular blood flow. The technique amounts to autocorrelation analysis of coherent photons after their migration through moving scatterers and subsequent collection by single-mode optical fibers. A primary cost driver of DCS instruments are the commercial hardware-based correlators, limiting the proliferation of multi-channel instruments for validation of perfusion analysis as a clinical diagnostic metric. We present the development of a low-cost scalable correlator enabled by microchip-based time-tagging, and a software-based multi-tau data analysis method. We will discuss the capabilities of the instrument as well as the implementation and validation of 2- and 8-channel systems built for live animal and pre-clinical settings.
Stable, relative localization of source and detection fibers is necessary for clinical implementation of quantitative optical perfusion monitoring methods such as diffuse correlation spectroscopy (DCS) and diffuse reflectance spectroscopy (DRS). A flexible and compact device design is presented as a platform for simultaneous monitoring of perfusion at a range of depths, enabled by precise location of optical fibers in a robust and secure adhesive patch. We will discuss preliminary data collected on human subjects in a lower body negative pressure model for hypovolemic shock. These data indicate that this method facilitates simple and stable simultaneous monitoring of perfusion at multiple depths and within multiple physiological compartments.
The vascularization and resulting perfusion of transferred tissues are critical to the success of grafts in buried free flap transplantations. To enable long-term clinical monitoring of grafted tissue perfusion during neovascularization and endothelialization, we are developing an implantable instrument for the continuous monitoring of perfusion using diffuse correlation spectroscopy (DCS), and augmented with diffuse reflectance spectroscopy (DRS). This work discusses instrument construction, integration, and preliminary results using a porcine graft model.
Dynamic maskless holographic lithography (DMHL) is a new micro-manufacturing technique that has no moving
parts. The laser light used for patterning is directed in all three dimensions with a hologram displayed on a
liquid-crystal spatial light modulator (SLM). Optical aberrations, like spherical aberration due to refractive index
mismatch between the photoresist and the immersion oil of the high-NA objective or astigmatism due to the
deformations in the surface of the SLM, can degrade the performance of the system. Degraded performance
includes a decrease in potential patterning volume and pattern fidelity and an increase in patterning time.
This paper presents a way to correct for these aberrations using Zernike polynomials. The optimal Zernike
coefficients are found by maximizing a sharpness metric. The effect of aberration correction on the DMHL process
is quantified by measuring the patterning volume. DMHL manufactured features made with this aberration
correction method show a marked improvement over features made without correction. It is even possible to
correct for misaligned optics with this method.
Dynamic maskless holographic lithography (DMHL) is a new micro-manufacturing technique that uses holograms
to create patterns on a substrate instead of a mask. In DMHL, gratings and Fresnel lenses are displayed on
nematic liquid crystal spatial light modulators (SLMs) to steer light to desired locations to expose sensitive
photopolymers. Micro-manufacturing can be done in two modes, serial or parallel. Serial refers to a beam being
scanned through a set of points and parallel refers to an entire intensity pattern being created at once. The
field over which patterning can be performed is affected by the diffraction efficiency of the displayed hologram,
the maximum possible spatial frequency of the SLM, and aliasing (light being steered to unintended spots due
to mismatches between designed and displayed phase patterns). This paper presents a technique to compensate
for these inherent inefficiencies by properly adjusting the amount of time spent by the beam at each point
in the desired feature, the dwell-time, during the lithographic process. The relationship between the spatial
frequency of the appropriate grating or Fresnel lens and the dwell time is discussed. Experiments are presented
with and without this technique applied, and results show that feature uniformity is improved with dwell-time
compensation.
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