Optical signature of nerve tissue-Exploratory ex vivo study comparing optical, histological, and molecular characteristics of different adipose and nerve tissues.

BACKGROUND: During several anesthesiological procedures, needles are inserted through the skin of a patient to target nerves. In most cases, the needle traverses several tissues-skin, subcutaneous adipose tissue, muscles, nerves, and blood vessels-to reach the target nerve. A clear identification of the target nerve can improve the success of the nerve block and reduce the rate of complications. This may be accomplished with diffuse reflectance spectroscopy (DRS) which can provide a quantitative measure of the tissue composition. The goal of the current study was to further explore the morphological, biological, chemical, and optical characteristics of the tissues encountered during needle insertion to improve future DRS classification algorithms. METHODS: To compare characteristics of nerve tissue (sciatic nerve) and adipose tissues, the following techniques were used: histology, DRS, absorption spectrophotometry, high-resolution magic-angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy, and solution 2D 13 C-1 H heteronuclear single-quantum coherence spectroscopy. Tissues from five human freshly frozen cadavers were examined. RESULTS: Histology clearly highlights a higher density of cellular nuclei, collagen, and cytoplasm in fascicular nerve tissue (IFAS). IFAS showed lower absorption of light around 1200 nm and 1750 nm, higher absorption around 1500 nm and 2000 nm, and a shift in the peak observed around 1000 nm. DRS measurements showed a higher water percentage and collagen concentration in IFAS and a lower fat percentage compared to all other tissues. The scattering parameter (b) was highest in IFAS. The HR-MAS NMR data showed three extra chemical peak shifts in IFAS tissue. CONCLUSION: Collagen, water, and cellular nuclei concentration are clearly different between nerve fascicular tissue and other adipose tissue and explain some of the differences observed in the optical absorption, DRS, and HR-NMR spectra of these tissues. Some differences observed between fascicular nerve tissue and adipose tissues cannot yet be explained but may be helpful in improving the discriminatory capabilities of DRS in anesthesiology procedures. Lasers Surg. Med. 50:948-960, 2018. © 2018 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Nerve detection using optical spectroscopy, an evaluation in four different models: In human and swine, in-vivo, and post mortem.

OBJECTIVE: Identification of peripheral nerve tissue is crucial in both surgery and regional anesthesia. Recently, optical tissue identification methods are presented to facilitate nerve identification in transcutaneous procedures and surgery. Optimization and validation of such techniques require large datasets. The use of alternative models to human in vivo, like human post mortem, or swine may be suitable to test, optimize and validate new optical techniques. However, differences in tissue characteristics and thus optical properties, like oxygen saturation and tissue perfusion are to be expected. This requires a structured comparison between the models. STUDY DESIGN: Comparative observational study. METHODS: Nerve and surrounding tissues in human (in vivo and post mortem) and swine (in vivo and post mortem) were structurally compared macroscopically, histologically, and spectroscopically. Diffuse reflective spectra were acquired (400-1,600 nm) after illumination with a broad band halogen light. An analytical model was used to quantify optical parameters including concentrations of optical absorbers. RESULTS: Several differences were found histologically and in the optical parameters. Histologically nerve and adipose tissue (subcutaneous fat and sliding fat) showed clear similarities between human and swine while human muscle enclosed more adipocytes and endomysial collagen. Optical parameters revealed model dependent differences in concentrations of ß-carotene, water, fat, and oxygen saturation. The similarity between optical parameters is, however, sufficient to yield a strong positive correlation after cross model classification. CONCLUSION: This study shows and discusses similarities and differences in nerve and surrounding tissues between human in vivo and post mortem, and swine in vivo and post mortem; this could support the discussion to use an alternative model to optimize and validate optical techniques for clinical nerve identification. Lasers Surg. Med. 50:253-261, 2018. © 2017 Wiley Periodicals, Inc.

Spectral tissue sensing to identify intra- and extravascular needle placement - A randomized single-blind controlled trial.

Safe vascular access is a prerequisite for intravenous drug admission. Discrimination between intra- and extravascular needle position is essential for procedure safety. Spectral tissue sensing (STS), based on optical spectroscopy, can provide tissue information directly from the needle tip. The primary objective of the trial was to investigate if STS can reliably discriminate intra-vascular (venous) from non-vascular punctures. In 20 healthy volunteers, a needle with an STS stylet was inserted, and measurements were performed for two intended locations: the first was subcutaneous, while the second location was randomly selected as either subcutaneous or intravenous. The needle position was assessed using ultrasound (US) and aspiration. The operators who collected the data from the spectral device were blinded to the insertion and ultrasonographic visualization procedure and the physician was blinded to the spectral data. Following offline spectral analysis, a prediction of intravascular or subcutaneous needle placement was made and compared with the "true" needle tip position as indicated by US and aspiration. Data for 19 volunteers were included in the analysis. Six out of 8 intended vascular needle placements were defined as intravascular according to US and aspiration. The remaining two intended vascular needle placements were negative for aspiration. For the other 11 final needle locations, the needle was clearly subcutaneous according to US examination and no blood was aspirated. The Mann-Whitney U test yielded a p-value of 0.012 for the between-group comparison. The differences between extra- and intravascular were in the within-group comparison computed with the Wilcoxon signed-rank test was a p-value of 0.022. In conclusion, STS is a promising method for discriminating between intravascular and extravascular needle placement. The information provided by this method may complement current methods for detecting an intravascular needle position.

The Role of Spectral Tissue Sensing During Lumbar Transforaminal Epidural Injection.

Spectral tissue sensing (STS) exploits the scattering and absorption of light by tissue. The main objective of the present study was to determine whether STS can discriminate between correct and incorrect placement of the needle tip during lumbar transforaminal epidural injection. This was a single-blind prospective observational study in 30 patients with lumbar radicular pain scheduled for lumbar transforaminal epidural injection. Spectral tissue sensing data from the needle tip were acquired along the needle trajectory at 4 predefined measurement points and compared with ultrasound, fluoroscopy, and digital subtraction angiography images. Spectral tissue sensing data contained the full spectra. The lipid and hemoglobin content at the different measurement points was also calculated, and partial least-squares discriminant analysis was used to estimate the sensitivity and specificity of STS. Spectral tissue sensing identified correct needle placement with a sensitivity of 57% and a specificity of 82%, and intraforaminal versus extraforaminal locations were identified with a sensitivity of 80% and a specificity of 71%.

Nerve detection with optical spectroscopy for regional anesthesia procedures.

BACKGROUND: Regional anesthesia has several advantages over general anesthesia but requires accurate needle placement to be effective. To achieve accurate placement, a needle equipped with optical fibers that allows tissue discrimination at the needle tip based on optical spectroscopy is proposed. This study investigates the sensitivity and specificity with which this optical needle can discriminate nerves from the surrounding tissues making use of different classification methods. METHODS: Diffuse reflectance spectra were acquired from 1563 different locations from 19 human cadavers in the wavelength range of 400-1710 nm; measured tissue types included fascicular tissue of the nerve, muscle, sliding fat and subcutaneous fat. Physiological parameters of the tissues were derived from the measured spectra and part of the data was directly compared to histology. Various classification methods were then applied to the derived parameter dataset to determine the accuracy with which fascicular tissue of the nerve can be discriminated from the surrounding tissues. RESULTS: From the parameters determined from the measured spectra of the various tissues surrounding the nerve, fat content, blood content, beta-carotene content and scattering were most distinctive when comparing fascicular and non-fascicular tissue. Support Vector Machine classification with a combination of feature selections performed best in discriminating fascicular nerve tissue from the surrounding tissues with a sensitivity and specificity around 90 %. CONCLUSIONS: This study showed that spectral tissue sensing, based on diffuse reflectance spectroscopy at the needle tip, is a promising technique to discriminate fascicular tissue of the nerve from the surrounding tissues. The technique may therefore improve accurate needle placement near the nerve which is necessary for effective nerve blocks in regional anesthesia.

Estimation of detection limits of a clinical fluorescence optical mammography system for the near-infrared fluorophore IRDye800CW: phantom experiments.

To evaluate if clinical fluorescence imaging of IRDye800CW is feasible on our fluorescence optical mammography system by estimating detection limits assessed by breast-cancer-simulating phantom experiments. Phantoms (2.1 cm(3), 0.9 cm(3)) with IRDye800CW concentrations of 0.5 to 120 nM were suspended in a 550 cm(3) measurement cup containing 507 surface-mounted source and detector fibers. The cup was filled with optical matching fluid containing IRDye800CW concentrations of 0, 5, 10, or 20 nM. Tomographic fluorescence images were acquired by exciting IRDye800CW at 730 nm; wavelengths above 750 nm were filtered. Signal intensities were calculated over a volume of interest corresponding to the size and location of the phantom in the reconstructed images. Correlations (R(2)) were calculated, and detection limits with associated upper 95% prediction interval were estimated. Between-day reproducibility was assessed with intraclass correlation coefficients (ICC). Fluorescent intensities were strongly correlated with phantom IRDye800CW concentrations (R(2)0.983 to 0.999). IRDye800CW detection limits ranged from 0.14 to 2.46 nM (upper 95% prediction limit 4.63 to 18.63 nM). ICC ranged from 0.88 to 1.00. The estimated detection limits for IRDye800CW were in the low-nanomolar range. These results support the start of clinical trials to evaluate the fluorescence optical mammography system using IRDye800CW labeled breast cancer targeting ligands.

Optical detection of peripheral nerves: an in vivo human study.

BACKGROUND AND OBJECTIVES: A critical challenge encountered in interventional pain medicine procedures is to accurately and efficiently identify transitions to peripheral nerve targets. Current methods, which include ultrasound guidance and nerve stimulation, are not perfect. In this pilot study, we investigated the feasibility of identifying tissue transitions encountered during insertions toward peripheral nerve targets using optical spectroscopy. METHODS: Using a custom needle stylet with integrated optical fibers, ultrasound-guided insertions toward peripheral nerves were performed in 20 patients, with the stylet positioned in the cannula of a 20-gauge stimulation needle. Six different peripheral nerves were represented in the study, with 1 insertion per patient. During each insertion, optical reflectance spectra were acquired with the needle tip in subcutaneous fat, skeletal muscle, and at the nerve target region. Differences in the spectra were quantified with 2 parameters that provide contrast for lipid and hemoglobin, respectively. RESULTS: The transition of the needle tip from subcutaneous fat to muscle was associated with lower lipid parameter values (P = 0.003) and higher hemoglobin parameter values (P = 0.023). The transition of the needle tip from the muscle to the nerve target region was associated with higher lipid parameter values (P = 0.008). CONCLUSIONS: The results indicate that the spectroscopic information provided by the needle stylet could potentially allow for reliable identification of transitions from subcutaneous fat to skeletal muscle and from the muscle to the nerve target region during peripheral nerve blocks.

Optical detection of vascular penetration during nerve blocks: an in vivo human study.

BACKGROUND AND OBJECTIVES: Complications resulting from vascular penetration during nerve blocks are rare but potentially devastating events that can occur despite meticulous technique. In this in vivo human pilot study, we investigated the potential for detecting vascular penetration with optical reflectance spectroscopy during blocks of the sympathetic chain and the communicating ramus at lumbar levels. METHODS: A custom-designed needle stylet with integrated optical fibers was used in combination with a commercial needle shaft. The needle stylet was connected to a console that delivered broadband light to tissue and spectrally resolved light that was scattered near the stylet tip. A total of 18 insertions were performed on 10 patients; testing for vascular penetration at the nerve target region was performed with aspiration and with radio-opaque contrast injections, visualized fluoroscopically. Optical absorption by hemoglobin was quantified with a blood parameter that was calculated from each spectrum. The blood parameter provided a measure of the difference between spectra acquired from the nerve target region and reference spectra acquired from blood extracted from a volunteer. RESULTS: In 2 insertions, vascular penetration was detected. Pronounced optical absorption by hemoglobin was observed to be associated with both of these events and absent in all other cases. The difference between the blood parameters obtained when vascular penetration was detected, and all other blood parameters were statistically significant (P = 0.006), with a diagnostic odds ratio of 35.4 (confidence interval, 2.21 to ∞). CONCLUSIONS: The results from this study suggest that optical spectroscopy has the potential to detect intravascular needle placement, which may in turn increase the safety of nerve blocks.

Diagnosis of breast cancer using diffuse optical spectroscopy from 500 to 1600 nm: comparison of classification methods.

We report on the use of diffuse optical spectroscopy analysis of breast spectra acquired in the wavelength range from 500 to 1600 nm with a fiber optic probe. A total of 102 ex vivo samples of five different breast tissue types, namely adipose, glandular, fibroadenoma, invasive carcinoma, and ductal carcinoma in situ from 52 patients were measured. A model deriving from the diffusion theory was applied to the measured spectra in order to extract clinically relevant parameters such as blood, water, lipid, and collagen volume fractions, ß-carotene concentration, average vessels radius, reduced scattering amplitude, Mie slope, and Mie-to-total scattering fraction. Based on a classification and regression tree algorithm applied to the derived parameters, a sensitivity-specificity of 98%-99%, 84%-95%, 81%-98%, 91%-95%, and 83%-99% were obtained for discrimination of adipose, glandular, fibroadenoma, invasive carcinoma, and ductal carcinoma in situ, respectively; and a multiple classes overall diagnostic performance of 94%. Sensitivity-specificity values obtained for discriminating malignant from nonmalignant tissue were compared to existing reported studies by applying the different classification methods that were used in each of these studies. Furthermore, in these reported studies, either lipid or ß-carotene was considered as adipose tissue precursors. We estimate both chromophore concentrations and demonstrate that lipid is a better discriminator for adipose tissue than ß-carotene.

Needle stylet with integrated optical fibers for spectroscopic contrast during peripheral nerve blocks.

The effectiveness of peripheral nerve blocks is highly dependent on the accuracy at which the needle tip is navigated to the target injection site. Even when electrical stimulation is utilized in combination with ultrasound guidance, determining the proximity of the needle tip to the target region close to the nerve can be challenging. Optical reflectance spectroscopy could provide additional information about tissues that is complementary to these navigation methods. We demonstrate a novel needle stylet for acquiring spectra from tissue at the tip of a commercial 20-gauge needle. The stylet has integrated optical fibers that deliver broadband light to tissue and receive scattered light. Two spectrometers resolve the light that is received from tissue across the wavelength range of 500-1600 nm. In our pilot study, measurements are acquired from a postmortem dissection of the brachial plexus of a swine. Clear differences are observed between spectra acquired from nerves and those acquired from adjacent tissue structures. We conclude that spectra acquired with the stylet have the potential to increase the accuracy with which peripheral nerve blocks are performed.

Epidural needle with embedded optical fibers for spectroscopic differentiation of tissue: ex vivo feasibility study.

Epidural injection is commonly used to provide intraoperative anesthesia, postoperative and obstetric analgesia, and to treat acute radicular pain. Identification of the epidural space is typically carried out using the loss of resistance (LOR) technique, but the usefulness of this technique is limited by false LOR and the inability to reliably detect intravascular or subarachnoid needle placement. In this study, we present a novel epidural needle that allows for the acquisition of optical reflectance spectra from tissue close to the beveled surface. This needle has optical fibers embedded in the cannula that deliver and receive light. With two spectrometers, light received from tissue is resolved across the wavelength range of 500 to 1600 nm. To determine the feasibility of optical tissue differentiation, spectra were acquired from porcine tissues during a post mortem laminectomy. The spectra were processed with an algorithm that derives estimates of the hemoglobin and lipid concentrations. The results of this study suggest that the optical epidural needle has the potential to improve the accuracy of epidural space identification.

Optical detection of the brachial plexus for peripheral nerve blocks: an in vivo swine study.

BACKGROUND AND OBJECTIVES: Accurate identification of nerves is critical to ensure safe and effective delivery of regional anesthesia during peripheral nerve blocks. Nerve stimulation is commonly used, but it is not perfect. Even when nerve stimulation is performed in conjunction with ultrasound guidance, determining when the needle tip is at the nerve target region can be challenging. In this in vivo pilot study, we investigated whether close proximity to the brachial plexus and penetration of the axillary artery can be identified with optical reflectance spectroscopy, using a custom needle stylet with integrated optical fibers. METHODS: Ultrasound-guided insertions to place the needle tip near the brachial plexus at the axillary level were performed at multiple locations in 2 swine, with the stylet positioned in the cannula of a 20-gauge stimulation needle. During each insertion, optical reflectance spectra were acquired with the needle tip in skeletal muscle, at the surface of muscle fascia, and at the nerve target region; confirmation of the final needle position was provided by nerve stimulation. In addition, an insertion to the lumen of the axillary artery was performed in a third swine. Differences in the spectra were quantified with lipid and hemoglobin parameters that provide contrast for optical absorption by the respective chromophores. RESULTS: The transition of the needle tip from skeletal muscle to the nerve target region was associated with higher lipid parameter values (P < 0.001) and lower hemoglobin parameter values (P < 0.001). The transition of the needle tip from muscle fascia to the nerve target region was associated with higher lipid parameter values (P = 0.001). Intraluminal access of the axillary artery was associated with an elevated hemoglobin parameter. CONCLUSIONS: Spectroscopic information obtained with the optical needle is distinct from nerve stimulation and complementary to ultrasound imaging, and it could potentially allow for reliable identification of the injection site during peripheral nerve blocks.

Optical mammography combined with fluorescence imaging: lesion detection using scatterplots.

Using scatterplots of 2 or 3 parameters, diffuse optical tomography and fluorescence imaging are combined to improve detectability of breast lesions. Small or low contrast phantom-lesions that were missed in the optical and fluorescence images were detected in the scatterplots. In patient measurements, all tumors were visible and easily differentiated from artifacts and areolas in the scatterplots. The different rate of intake and wash out of the fluorescent contrast agent in the healthy versus malignant tissues was also observed in the scatterplot: this information can be used to discriminate malignant lesion from normal structures.

Effect of bile absorption coefficients on the estimation of liver tissue optical properties and related implications in discriminating healthy and tumorous samples.

We investigated differences between healthy tissue and metastatic tumor from ex vivo human partial liver resections using diffuse optical spectroscopy with a fiber optic probe. We extracted various physiological and morphological parameters from the spectra. During evaluation of the residual between the measurements and a fit model based on diffusion theory, we found that bile is an additional chromophore absorbing in the visible wavelength range that was missing in our model. Consistency of the residual with the absorption spectrum of bile was noticed. An accurate measurement of the absorption coefficient of bile from various human bile samples was performed and implemented into the fit model. Having the absorption coefficient of bile as a priori knowledge in the model showed a clear improvement in terms of reducing the fitting discrepancies. The addition of this chromophore yields significantly different estimates of the amount of blood. Furthermore, the estimated bile volume fraction and reduced scattering amplitude turned out to be two main relevant discriminators between normal and metastatic liver tissues.

Identification of the epidural space with optical spectroscopy: an in vivo swine study.

BACKGROUND: Accurate identification of the epidural space is critical for safe and effective epidural anesthesia or treatment of acute lumbar radicular pain with epidural steroid injections. The loss-of-resistance technique is commonly used, but it is known to be unreliable. Even when it is performed in conjunction with two-dimensional fluoroscopic guidance, determining when the needle tip enters the epidural space can be challenging. In this swine study, we investigated whether the epidural space can be identified with optical spectroscopy, using a custom needle with optical fibers integrated into the cannula. METHODS: Insertion of the needle tip into the epidural space was performed with midline and paramedian approaches in a swine. In each insertion, optical spectra were acquired at different insertion depths, and anatomical localization of the needle was determined by three-dimensional imaging with rotational C-arm computed tomography. Optical spectra that included both visible and near-infrared wavelength ranges were processed to derive estimates of the blood and lipid volume fractions. RESULTS: In all insertions, the transition of the needle tip to the epidural space from an adjacent tissue structure (interspinous ligament or the ligamentum flavum) was found to be associated with an increase in the lipid volume fraction. These increases, which ranged from 1.6- to 3.0-fold, were statistically significant (P = 0.0020). Lipid fractions obtained from the epidural space were 1.9- to 20-fold higher than those obtained from muscle (P = 0.0013). Accidental penetration of an epidural vein during one insertion coincided with a high blood volume fraction. CONCLUSIONS: The spectroscopic information obtained with the optical spinal needle is complementary to fluoroscopic images, and it could potentially allow for reliable identification of the epidural space during needle placement.

Estimation of lipid and water concentrations in scattering media with diffuse optical spectroscopy from 900 to 1,600 nm.

We demonstrate a method to estimate the concentrations of water and lipid in scattering media such as biological tissues with diffuse optical spectra acquired over the range of 900 to 1600 nm. Estimations were performed by fitting the spectra to a model of light propagation in scattering media derived from diffusion theory. To validate the method, spectra were acquired from tissue phantoms consisting of lipid and water emulsions and swine tissues ex vivo with a two-fiber probe.

Estimation of biological chromophores using diffuse optical spectroscopy: benefit of extending the UV-VIS wavelength range to include 1000 to 1600 nm.

With an optical fiber probe, we acquired spectra from swine tissue between 500 and 1600 nm by combining a silicon and an InGaAs spectrometer. The concentrations of the biological chromophores were estimated by fitting a mathematical model derived from diffusion theory. The advantage of our technique relative to those presented in previous studies is that we extended the commonly-used wavelength ranges of 500 and 1000 nm to include the range of 1000 to 1600 nm, where additional water and lipid absorption features exist. Hence, a more accurate estimation of these two chromophores is expected when spectra are fitted between 500 and 1600 nm than between 500 and 1000 nm. When extending the UV-VIS wavelength range, the estimated total amount of chromophores approached 100% of the total as present in the probed volume. The confidence levels of the water and lipid related parameters increases by a factor of four.

A novel fluorescent imaging agent for diffuse optical tomography of the breast: first clinical experience in patients.

PURPOSE: This is the first clinical evaluation of a novel fluorescent imaging agent (Omocianine) for breast cancer detection with diffuse optical tomography (DOT). PROCEDURES: Eleven women suspected of breast cancer were imaged with DOT at multiple time points (up to 24 h) after receiving an intravenous injection of Omocianine (doses 0.01 to 0.1 mg/kg bodyweight). Breast MRI was obtained for comparison. RESULTS: Histopathology showed invasive cancer in ten patients and fibroadenoma in one patient. With the lowest dose of Omocianine, two of three lesions were detected; with the second dose, three of three lesions were detected; with the two highest doses, none of five lesions were detected. Lesion location on DOT showed excellent agreement with MRI. Optimal lesion-to-background signals were obtained after 8 h. No adverse events occurred. CONCLUSIONS: Lowest doses of Omocianine performed best in lesion detection; DOT using a low-dose fluorescent agent is feasible and safe for breast cancer visualization in patients.

Diffuse optical tomography of the breast: preliminary findings of a new prototype and comparison with magnetic resonance imaging.

This paper presents an evaluation of a prototype diffuse optical tomography (DOT) system. Seventeen women with 18 breast lesions (10 invasive carcinomas, 2 fibroadenomas, and 6 benign cysts; diameters 13-54 mm) were evaluated with DOT and magnetic resonance imaging (MRI). A substantial fraction of the original 36 recruited patients could not be examined using this prototype due to technical problems. A region of interest (ROI) was drawn at the lesion position as derived from MRI and at the mirror image site in the contralateral healthy breast. ROIs were assessed quantitatively and qualitatively by two observers independently in two separate readings. Intra- and interobserver agreements were calculated using kappa statistics (k) and intraclass correlation coefficients (ICCs). Discriminatory values for presence of malignancy were determined by receiver operating characteristic (ROC) analyses. Intraobserver agreements were excellent (k 0.88 and 0.88; ICC 0.978 and 0.987), interobserver agreements were good to excellent (k 0.77-0.95; ICC 0.96-0.98). Discriminatory values for presence of malignancy were 0.92-0.93 and 0.97-0.99 for quantitative and qualitative ROC analysis, respectively. This DOT system has the potential to discriminate malignant from benign breast tissue in a reproducible qualitative and quantitative manner. Important technical improvements are required before this technique is ready for clinical application.

Diffuse optical tomography of the breast: initial validation in benign cysts.

PURPOSE: The purpose of this study was to validate a newly developed diffuse optical tomography (DOT) system on benign cysts in the breast. PROCEDURES: Eight patients with 20 benign cysts were included. Study procedures consisted of optical breast imaging and breast magnetic resonance imaging (MRI) for comparison. A reconstruction algorithm computed three-dimensional images for each of the four near-infrared wavelengths used by our DOT system (Philips Healthcare, Best, The Netherlands). These images were combined using a spectroscopic model to assess tissue composition and lesion size. RESULTS: Twenty cysts were analyzed in eight patients. By using the spectroscopic information, 13 of 20 cysts (65%) were visualized with DOT, confirming their high water and low total hemoglobin content. Lesion size and location showed good agreement with MRI; Pearson correlation coefficient was 0.7 (p < 0.01). CONCLUSIONS: DOT can visualize benign cysts in the breast and elucidate their high water and low total hemoglobin content by spectroscopic analysis.