Calibration and validation scheme for in vivo spectroscopic imaging of tissue oxygenation.

The determination of the level of oxygenation in optically accessible tissues using multispectral or hyperspectral imaging (HSI) of oxy- and deoxyhemoglobin has special appeal in clinical work due to its noninvasiveness, ease of use, and capability of providing molecular and anatomical information at near video rates during surgery. In this paper we refer to an example of the use of HSI in monitoring oxygenation of kidneys during partial nephrectomy. In a study using porcine models, it was found that artery-only clamping left the kidney better oxygenated, as opposed to simultaneously clamping the artery and the vein. A subsequent study correlates gradations in blood flow by partial clamping during the surgical procedure with postoperative renal function via assessment of creatinine level. We discuss the various contributions to the uncertainty of the oxygen saturation measured by this remote-sensing imaging technique in medical application.

Active DLP hyperspectral illumination: a noninvasive, in vivo, system characterization visualizing tissue oxygenation at near video rates.

We report use of a novel hyperspectral imaging system utilizing digital light processing (DLP) technology to noninvasively visualize in vivo tissue oxygenation during surgical procedures. The system's novelty resides in its method of illuminating tissue with precisely predetermined continuous complex spectra. The Texas Instruments digital micromirror device, DMD, chip consisting of 768 by 1024 mirrors, each 16 µm square, can be switched between two positions at 12.5 kHz. Switching the appropriate mirrors controls the intensity of light illuminating the tissue as a function of wavelength, active spectral illumination. Meaning, the tissue can be illuminated with a different spectrum of light within 80 µs. Precisely, predetermined spectral illumination penetrates into patient tissue, its chemical composition augments the spectral properties of the light, and its reflected spectra are detected and digitized at each pixel detector of a silicon charge-coupled device, CCD. Using complex spectral illumination, digital signal processing and chemometric methods produce chemically relevant images at near video rates. Specific to this work, tissue is illuminated spectrally with light spanning the visible electromagnetic spectrum (380 to 780 nm). Spectrophotometric images are detected and processed visualizing the percentage of oxyhemoglobin at each pixel detector and presented continuously, in real time, at 3 images per second. As a proof of principle application, kidneys of four live anesthetized pigs were imaged before, during, and after renal vascular occlusion. DLP Hyperspectral Imaging with active spectral illumination detected a 64.73 ± 1.5% drop in the oxygenation of hemoglobin within 30 s of renal arterial occlusion. Producing chemically encoded images at near video rate, time-resolved hyperspectral imaging facilitates monitoring renal blood flow during animal surgery and holds considerable promise for doing the same during human surgical interventions.

Assessment of renal oxygenation during partial nephrectomy using hyperspectral imaging.

PURPOSE: DLP® hyperspectral imaging is a technology that can be used to construct a highly sensitive, noninvasive, real-time tissue hemoglobin saturation map. This almost video rate technology may be a tool to monitor renal perfusion/oxygenation during hilar occlusion and kidney recovery. We describe our initial experience using hyperspectral imaging to assess renal hemoglobin saturation parameters during open partial nephrectomy for renal cortical tumors in humans. MATERIALS AND METHODS: Hyperspectral images were collected intraoperatively during open partial nephrectomy. The kidney was actively illuminated using a hyperspectral imaging camera with visible light consisting of a chemometrically predetermined spectrum (520 to 645 nm) for hemoglobin. Spectroscopic reflectance images were captured by a focal plane array, which were digitally processed to visualize the percent of oxyhemoglobin at each image pixel. RESULTS: Hyperspectral imaging was done in 21 patients with a mean age of 56 years who were undergoing partial nephrectomy. Mean clamp time was 37.0 minutes. Median baseline percent of oxyhemoglobin in all patients was 74.6%. Hyperspectral imaging revealed a median 20.0% decrease from normalized pre-occlusion baseline at a median 10.3 minutes of hilar occlusion, where it plateaued for the duration of kidney ischemia. Upon reperfusion the percent of oxyhemoglobin returned to baseline at a median of 5.8 minutes. CONCLUSIONS: Hyperspectral imaging is a real-time noninvasive method to assess renal oxyhemoglobin saturation intraoperatively throughout the kidney. A nadir percent of oxyhemoglobin is attained within 10 minutes of hilar occlusion. This knowledge may allow future surgical or pharmacological interventions that titrate or minimize ischemic injury in real time.

Renal oxygenation measurement during partial nephrectomy using hyperspectral imaging may predict acute postoperative renal function.

BACKGROUND AND PURPOSE: Digital Light Processing hyperspectral imaging (HsI) produces a highly sensitive, real-time tissue oxygenation map to monitor renal perfusion/oxygenation during partial nephrectomy (PN). Our initial experience with HsI revealed considerable variation in the baseline renal oxygenation, and we sought to correlate these differences with postoperative renal function. PATIENTS AND METHODS: Hyperspectral images were collected intraoperatively in patients undergoing PN for cortical tumors. The kidney was illuminated with visible light (520-645 nm), the spectrum corresponding to that of oxyhemoglobin. Reflectance images were captured and digitally processed to determine the percentage of oxyhemoglobin (HbO2) at each image pixel. Images were taken before hilar occlusion; these %HbO2 values were used to categorize patients as high (>75% HbO2) or low (<75% HbO2) oxygenation, and postoperative eGFR was assessed. RESULTS: There were 26 patients who underwent PN with ice cooling and HsI imaging. Nineteen patients had a "low" preclamp oxygenation (mean 69%) while the other 7 had a "high" HbO2 (mean 77%). There was no difference in tumor size, hematocrit value, clamp time, or preoperative eGFR between the two groups. Patients with a higher baseline %HbO2 had no significant postoperative change in their eGFR (mean 0 mL/min/1.73 m(2), +4%), while those with the lower baseline %HbO2 had a significant acute decline (mean 15 mL/min/1.73 m(2), -20%, P=0.02, 0.006). CONCLUSIONS: Baseline renal oxygenation, as measured with HsI, may help predict risk of postoperative renal insufficiency and may distinguish between patients with otherwise similar baseline characteristics, such as eGFR. HsI may provide individualized assessment of renal function to influence intraoperative decision-making to help preserve renal function.

Renal oxygenation during robot-assisted laparoscopic partial nephrectomy: characterization using laparoscopic digital light processing hyperspectral imaging.

UNLABELLED: Abstract Background and Purpose: Digital light processing-based hyperspectral imaging (DLP(®)-HSI) was adapted for use during laparoscopic surgery by coupling the spectral illumination source with a conventional laparoscopic light guide and incorporating a customized digital charge-coupled device camera for image acquisition. The system was used to characterize renal oxygenation during robot-assisted laparoscopic partial nephrectomy (RALPN) in humans. PATIENTS AND METHODS: After Institutional Review Board approval, laparoscopic DLP-HSI was performed in consecutive patients undergoing RALPN at our institution. Time trends in relative tissue oxygen saturation (%HbO2) were descriptively analyzed. Associations between %HbO2 and patient age, comorbidities, and estimated glomerular filtration rate (eGFR) were investigated using the Kendall tau test. RESULTS: Laparoscopic DLP-HSI was performed in 18 patients between May 2011 and February 2012. Median (interquartile range; IQR) age was 55.9 (49-67.5) years. Of the patients, 10/18 (56%) were men and 12/18 (66.7%) had a history of hypertension, diabetes, and/or tobacco use. Median (IQR) %HbO2 before, during, and after ischemia was 60.8% (57.9-68.2%), 53.6% (46.8-55.1%), and 61.5% (54.9-67.6%), respectively. Baseline %HbO2 was inversely associated with preoperative eGFR (τ=-0.38; P=0.036), and eGFR at most recent follow-up (τ=-0.38; P=0.036). Baseline or ischemic %HbO2 did not correlate with hypertension, diabetes, and/or tobacco history. Younger patients (<56 years) had a lower median baseline %HbO2 (P=0.07) and a higher median preoperative eGFR (P=0.038), than their older counterparts. CONCLUSION: The laparoscopic HSI system successfully characterized dynamic changes in renal oxygenation during RALPN. Intraoperative laparoscopic HSI outcomes have the potential to predict postoperative individual kidney function.

Minimal arterial in-flow protects renal oxygenation and function during porcine partial nephrectomy: confirmation by hyperspectral imaging.

OBJECTIVES: To examine the potential for renal protection through incomplete renal artery (RA) occlusion with both assessments of creatinine changes and the use of hyperspectral imaging to monitor tissue oxygenation. Renal ischemia during partial nephrectomy can have adverse consequences on renal function. METHODS: Fourteen pigs with a solitary kidney underwent open partial nephrectomy with warm ischemia. The RA flow was measured and reduced to 25%, 10%, and 0% of baseline for 60 minutes. Hyperspectral imaging was used to assess the percentage of oxyhemoglobin (%HbO(2)) at baseline, during ischemia, and during reperfusion. The %HbO(2) and change in the serum creatinine level from baseline were compared. RESULTS: The baseline RA flow and %HbO(2) were similar in all groups, and, as expected, RA occlusion resulted in decreasing %HbO(2). The reduction of RA flow to 25% and 10% improved the nadir tissue oxygenation compared with 0% flow (P = .01 and P = .04, respectively) and 25% flow also appeared to prolong the interval to reach the nadir %HbO(2). Reperfusion resulted in a swift return to the baseline %HbO(2) in all 3 groups. The change in the serum creatinine from baseline to postoperative day 7 showed significantly improved renal preservation in the 25% RA flow group. CONCLUSIONS: Incomplete RA occlusion during porcine partial nephrectomy resulted in favorable renal oxygenation profiles with as little as 10% blood flow and appeared to be renoprotective when 25% of the baseline RA flow is preserved. Hyperspectral imaging is a sensitive, noninvasive tool for real-time monitoring of renal oxygenation and, thereby, blood flow, which could facilitate intraoperative decision-making to protect kidney function.

Characterization of renal ischemia using DLP hyperspectral imaging: a pilot study comparing artery-only occlusion versus artery and vein occlusion.

BACKGROUND AND PURPOSE: Renal artery-only (AO) occlusion, as opposed to artery and vein (AV) occlusion, has demonstrated some benefit in reducing renal insufficiency during warm ischemia. In this pilot study, we used digital light projection hyperspectral imaging (HSI) to construct a "real time" tissue oxygenation "map" to determine whether there are differences in renal tissue oxygenation during vascular occlusion with AO vs AV. MATERIALS AND METHODS: Renal vascular occlusion with either AO or AV was performed for 60 minutes in seven porcine renal units. Using HSI, the percentage of oxyhemoglobin (%HbO(2)) in the renal cortex was determined at 4-minute increments throughout the ischemic period and for 30 minutes after reperfusion. RESULTS: Average baseline %HbO(2) in all animals was approximately 70%. After vascular occlusion in both cohorts, %HbO(2) decreased by one third within 2 to 5 minutes, with a gradual decline in %HbO(2) over the remaining 55 minutes. Oxyhemoglobin profiles for AO and AV occlusion diverged significantly between 16 and 24 minutes after vascular occlusion (P = 0.0001 and 0.036, respectively), with a merging of the two curves occurring after approximately 36 minutes (P = 0.093). During reperfusion, average %HbO(2) improved to 72.4% after 25 to 30 minutes. CONCLUSION: In this pilot study, we demonstrate that renal tissue oxygenation drops rapidly after occlusion of the renal vasculature and returns to near baseline 30 minutes after reperfusion. In the porcine model, the %HbO(2) differs significantly between AO and AV occlusion for up to 35 minutes after ischemia onset, indicating a possible "ischemic window" in which AO occlusion may provide benefit over AV occlusion.

Intraoperative bile duct visualization using near-infrared hyperspectral video imaging.

BACKGROUND: Current methodologies for imaging the biliary system during cholecystectomy are cumbersome and do not eliminate the risk of bile duct injury. We describe an approach to intraoperative biliary imaging that will enable surgeons to see through the hepatoduodenal ligament and visualize the anteriorly placed biliary system. METHODS: A laparoscopic-capable, near-infrared, hyperspectral imaging system was built. Reflected light passes through a liquid crystal filter that is continuously tunable in the near-infrared spectrum (650-1,100 nm). Spectroscopic image data are collected from laparoscopic surgery images onto array detectors formatted into a 3-dimensional hyperspectral data cube having spatially resolved images in the x-y plane and wavelength data in the z plane. Deconvoluting and color-coding the spatial and spectral information provides an image representative of inherent chemical properties to the imaged tissue. RESULTS: Images of porcine biliary structures were obtained. The common duct-reflected spectra displayed a characteristic lipid shoulder at 930 nm and a strong water peak at 970 nm. Venous structures had absorption peaks at 760 nm (deoxyhemoglobin), 800 nm (oxyhemoglobin), and 970 nm (water). Arterial vessels had absorption peaks at 800 nm and 970 nm that would be expected for oxyhemoglobin and water. CONCLUSIONS: We have designed and constructed a device to significantly enhance intraoperative biliary imaging. This system should enable surgeons to see through the hepatoduodenal ligament and image the anteriorly placed biliary system without the need for dissection of the cystic duct, as is needed with intraoperative cholangiography. Because the biliary system can be seen before any dissection is performed, this dimensional imaging technology has the potential for eradicating bile duct injury.

Characterization of a near-infrared laparoscopic hyperspectral imaging system for minimally invasive surgery.

We developed and characterized a new imaging platform for minimally invasive surgical venues, specifically a system to help guide laparoscopic surgeons to visualize biliary anatomy. This platform is a novel combination of a near-infrared hyperspectral imaging system coupled with a conventional surgical laparoscope. Intraoperative tissues are illuminated by optical fibers arranged in a ring around a center-mounted relay lens collecting back-reflected light from tissues to the hyperspectral imaging system. The system consists of a focal plane array (FPA) and a liquid crystal tunable filter, which is continuously tunable in the near-infrared spectral range of 650-1100 nm with the capability of passing light with a mean bandwidth of 6.95 nm, and the FPA is a high-sensitivity back-illuminated, deep depleted charge-coupled device. Placing a standard resolution target 5.1 cm from the distal end of the laparoscope, a typical intraoperative working distance, produced a 7.6-cm-diameter field of view with an optimal spatial resolution of 0.24 mm. In addition, the system's spatial and spectral resolution and its wavelength tuning accuracy are characterized. The spectroscopic images are formatted into a three-dimensional hyperspectral image cube and processed using principle component analysis. The processed images provide contrast based on measured spectra associated with chemically different anatomical structures helping identify the main molecular chromophores inherent to each tissue. The principal component images were found to image swine gallbladder and biliary structures from surrounding tissues, in real time, during cholecystectomy surgery. Furthermore, it is shown that surgeons can interrogate selected image subregions for their molecular composition identifying biliary anatomy during surgery and before any invasive action is undertaken.

Imaging hemoglobin oxygen saturation in sickle cell disease patients using noninvasive visible reflectance hyperspectral techniques: effects of nitric oxide.

Sickle cell disease is characterized by microvascular occlusion and hemolytic anemia, factors that impair tissue oxygen delivery. We use visible reflectance hyperspectral imaging to quantitate skin tissue hemoglobin oxygen saturation (HbO2) and to determine whether changes in blood flow during nitric oxide (NO) stimulation or gas administration (therapies proposed for this disease) improve skin tissue oxygen saturation in five patients with sickle cell disease. Compared with six healthy African-American subjects, sickle cell patients exhibited higher forearm blood flows (7.4 +/- 1.8 vs. 3.2 +/- 0.4 ml.min-1.100 ml tissue-1, P = 0.037) but significantly reduced percentages of skin HbO2 (61.0 +/- 0.2 vs. 77.5 +/- 0.2%, P < 0.001). Administration of acetylcholine to patients increased blood flow by 15.1 +/- 3.8 ml.min-1.100 ml tissue-1 and the percentage of skin HbO2 by 4.1 +/- 0.3% (P = 0.02, P < 0.001, respectively, from baseline values). Sodium nitroprusside, a direct NO donor, increased blood flow by 3.9 +/- 1.1 ml/min and the percentage of skin HbO2 by 2.9 +/- 0.3% (P = 0.02, P < 0.001, respectively). NO inhalation had no effect on forearm blood flow, yet increased the percentage of skin HbO2 by 2.3 +/- 0.3% (P < 0.001). Percentages of skin HbO2 were exponentially related to blood flow (R = 0.97, P < 0.001), indicating a limit to skin tissue oxygen saturation at high blood flows. Thus, for acetylcholine infusion leading to blood flows sevenfold greater than those of healthy resting African-American subjects, patients still exhibited lower percentages of skin HbO2 (65.2 +/- 0.2 vs. 77.5 +/- 0.2%, P < 0.001). Visible reflectance hyperspectral imaging demonstrates that either the stimulation or the administration of NO pharmacologically or by gas inhalation improves, but does not normalize, skin tissue oxygen saturation in patients with sickle cell disease.

Visible reflectance hyperspectral imaging: characterization of a noninvasive, in vivo system for determining tissue perfusion.

We characterize a visible reflectance hyperspectral imaging system for noninvasive, in vivo, quantitative analysis of human tissue in a clinical environment. The subject area is illuminated with a quartz-tungsten-halogen light source, and the reflected light is spectrally discriminated by a liquid crystal tunable filter (LCTF) and imaged onto a silicon charge-coupled device detector. The LCTF is continuously tunable within its useful visible spectral range (525-725 nm) with an average spectral full width at half-height bandwidth of 0.38 nm and an average transmittance of 10.0%. A standard resolution target placed 5.5 ft from the system results in a field of view with a 17-cm diameter and an optimal spatial resolution of 0.45 mm. The measured reflectance spectra are quantified in terms of apparent absorbance and formatted as a hyperspectral image cube. As a clinical example, we examine a model of vascular dysfunction involving both ischemia and reactive hyperemia during tissue reperfusion. In this model, spectral images, based upon oxyhemoglobin and deoxyhemoblobin signals in the 525-645-nm region, are deconvoluted using a multivariate least-squares regression analysis to visualize the spatial distribution of the percentages of oxyhemoglobin and deoxyhemoglobin in specific skin tissue areas.