Raman spectroscopy in microsurgery: impact of operating microscope illumination sources on data quality and tissue classification.

Ambient light artifacts can confound Raman spectroscopy measurements performed in a clinical setting such as during open surgery. However, requiring light sources to be turned off during intraoperative spectral acquisition can be impractical because it can slow down the procedure by requiring surgeons to acquire data under light conditions different from the routine clinical practice. Here a filter system is introduced allowing in vivo Raman spectroscopy measurements to be performed with the light source of a neurosurgical microscope turned on, without interfering with the standard procedure. Ex vivo and in vivo results on calf and human brain, respectively, show that when the new filter system is used there is no significant difference between Raman spectra acquired under pitch dark conditions or with the microscope light source turned on. This is important for the clinical translation of Raman spectroscopy because of the resulting decrease in total imaging time for each measurement and because the surgeon can now acquire spectroscopic data with no disruption of the surgical workflow.

Sub-diffuse interstitial optical tomography to improve the safety of brain needle biopsies: a proof-of-concept study.

The extraction of tissue samples during brain needle biopsy can cause life-threatening hemorrhage because of significant blood vessel injury during the procedure. Vessel rupture can have significant consequences for patient health, ranging from transient neurological deficits to death. Here, we present a sub-diffuse optical tomography technique that can be integrated into neurosurgical workflow to detect the presence of blood vessels. A proof-of-concept study performed on a realistic brain tissue phantom is presented and demonstrates that interstitial optical tomography (iOT) can detect several 1 mm diameter high-contrast absorbing objects located <2 mm from the needle.

Multispectral intraoperative imaging for the detection of the hemodynamic response to interictal epileptiform discharges.

Interictal epileptiform discharges (IEDs) are brief neuronal discharges occurring between seizures in patients with epilepsy. The characterization of the hemodynamic response function (HRF) specific to IEDs could increase the accuracy of other functional imaging techniques to localize epileptiform activity, including functional near-infrared spectroscopy and functional magnetic resonance imaging. This study evaluated the possibility of using an intraoperative multispectral imaging system combined with electrocorticography (ECoG) to measure the average HRF associated with IEDs in eight patients. Inter-patient variability of the HRF is illustrated in terms of oxygenated hemoglobin peak latency, oxygenated hemoglobin increase/decrease following IEDs, and signal-to-noise ratio. A sub-region was identified using an unsupervised clustering algorithm in three patients that corresponded to the most active area identified by ECoG.

Multispectral diffuse reflectance can discriminate blood vessels and bleeding during neurosurgery based on low-frequency hemodynamics.

SIGNIFICANCE: The practicality of optical methods detecting tissue optical contrast (absorption, elastic and inelastic scattering, fluorescence) for surgical guidance is limited by interferences from blood pooling and the resulting partial or complete inability to interrogate cortex and blood vessels. AIM: A multispectral diffuse reflectance technique was developed for intraoperative brain imaging of hemodynamic activity to automatically discriminate blood vessels, cortex, and bleeding at the brain surface. APPROACH: A manual segmentation of blood pooling, cortex, and vessels allowed the identification of a frequency range in hemoglobin concentration variations associated with high optical signal in blood vessels and cortex but not in bleeding. Reflectance spectra were then used to automatically segment areas with and without hemodynamic activity as well as to discriminate blood from cortical areas. RESULTS: The frequency range associated with low-frequency hemodynamics and respiratory rate (0.03 to 0.3 Hz) exhibits the largest differences in signal amplitudes for bleeding, blood vessels, and cortex. A segmentation technique based on simulated reflectance spectra initially allowed discrimination of blood (bleeding and vessels) from cortical tissue. Then, a threshold applied to the low-frequency components from deoxyhemoglobin allowed the segmentation of bleeding from vessels. A study on the minimum acquisition time needed to discriminate all three components determined that ∼25 s was necessary to detect changes in the low-frequency range. Other frequency ranges such as heartbeat (1 to 1.7 Hz) can be used to reduce the acquisition time to few seconds but would necessitate optimizing instrumentation to ensure larger signal-to-noise ratios are achieved. CONCLUSIONS: A method based on multispectral reflectance signals and low-frequency hemoglobin concentration changes can be used to distinguish bleeding, blood vessels, and cortex. This could be integrated into fiber optic probes to enhance signal specificity by providing users an indication of whether measurements are corrupted by blood pooling, an important confounding factor in biomedical optics applied to surgery.

Wide-field optical spectroscopy system integrating reflectance and spatial frequency domain imaging to measure attenuation-corrected intrinsic tissue fluorescence in radical prostatectomy specimens.

The development of a multimodal optical imaging system is presented that integrates endogenous fluorescence and diffuse reflectance spectroscopy with single-wavelength spatial frequency domain imaging (SFDI) and surface profilometry. The system images specimens at visible wavelengths with a spatial resolution of 70 µm, a field of view of 25 cm2 and a depth of field of ∼1.5 cm. The results of phantom experiments are presented demonstrating the system retrieves absorption and reduced scattering coefficient maps using SFDI with <6% reconstruction errors. A phase-shifting profilometry technique is implemented and the resulting 3-D surface used to compute a geometric correction ensuring optical properties reconstruction errors are maintained to <6% in curved media with height variations <20 mm. Combining SFDI-computed optical properties with data from diffuse reflectance spectra is shown to correct fluorescence using a model based on light transport in tissue theory. The system is used to image a human prostate, demonstrating its ability to distinguish prostatic tissue (anterior stroma, hyperplasia, peripheral zone) from extra-prostatic tissue (urethra, ejaculatory ducts, peri-prostatic tissue). These techniques could be integrated in robotic-assisted surgical systems to enhance information provided to surgeons and improve procedural accuracy by minimizing the risk of damage to extra-prostatic tissue during radical prostatectomy procedures and eventually detect residual cancer.

Intraoperative video-rate hemodynamic response assessment in human cortex using snapshot hyperspectral optical imaging.

Using light, we are able to visualize the hemodynamic behavior of the brain to better understand neurovascular coupling and cerebral metabolism. In vivo optical imaging of tissue using endogenous chromophores necessitates spectroscopic detection to ensure molecular specificity as well as sufficiently high imaging speed and signal-to-noise ratio, to allow dynamic physiological changes to be captured, isolated, and used as surrogate of pathophysiological processes. An optical imaging system is introduced using a 16-bands on-chip hyperspectral camera. Using this system, we show that up to three dyes can be imaged and quantified in a tissue phantom at video-rate through the optics of a surgical microscope. In vivo human patient data are presented demonstrating brain hemodynamic response can be measured intraoperatively with molecular specificity at high speed.