Hyperspectral imaging for monitoring of free flaps of the oral cavity: A feasibility study.

OBJECTIVES: Hyperspectral imaging (HSI) provides spectral information about hemoglobin, water and oxygen supply and has thus great potential in perfusion monitoring. The aim of the present study was to investigate the feasibility of HSI in the postoperative monitoring of intraoral free flaps. METHODS: The 14 patients receiving reconstructive head and neck surgery with a radial forearm free flap were included. HSI was performed intraoperatively (t0), on Day 1 (t1), 2 (t2), 3-6 (t3), 7-9 (t4), 10-11 (t5) and 12-15 (t6) postoperatively. Flap tissue perfusion was assessed on defined regions of interest by calculating the perfusion indices Tissue Hemoglobin Index (THI), hemoglobin oxygenation (StO2 ), Near Infrared Perfusion Index (NIR Perfusion Index) and Tissue Water Index (TWI). RESULTS: Image quality varied depending on location of the flap and time of measurement. StO2 was >50 intraoperatively and >40 on t1 for all patients. A significant difference was found solely for TWI between t0 and t2 and t0 and t4. No flap loss occurred. CONCLUSIONS: The use of HSI in the monitoring of intraoral flaps is feasible and might become a valuable addition to the current clinical examination of free flaps.

Comprehensive Investigation of Parameters Influencing Fluorescence Lifetime Imaging Microscopy in Frequency- and Time-Domain Illustrated by Phasor Plot Analysis.

Having access to fluorescence lifetime, researchers can reveal in-depth details about the microenvironment as well as the physico-chemical state of the molecule under investigation. However, the high number of influencing factors might be an explanation for the strongly deviating values of fluorescent lifetimes for the same fluorophore reported in the literature. This could be the reason for the impression that inconsistent results are obtained depending on which detection and excitation scheme is used. To clarify this controversy, the two most common techniques for measuring fluorescence lifetimes in the time-domain and in the frequency-domain were implemented in one single microscopy setup and applied to a variety of fluorophores under different environmental conditions such as pH-value, temperature, solvent polarity, etc., along with distinct state forms that depend, for example, on the concentration. From a vast amount of measurement results, both setup- and sample-dependent parameters were extracted and represented using a single display form, the phasor-plot. The measurements showed consistent results between the two techniques and revealed which of the tested parameters has the strongest influence on the fluorescence lifetime. In addition, quantitative guidance as to which technique is most suitable for which research task and how to perform the experiment properly to obtain consistent fluorescence lifetimes is discussed.

Development of a microstructured tissue phantom with adaptable optical properties for use with microscopes and fluorescence lifetime imaging systems.

OBJECTIVES: For the development and validation of diagnostic procedures based on microscopic methods, knowledge about the imaging depth and achievable resolution in tissue is crucial. This poses the challenge to develop a microscopic artificial phantom focused on the microscopic instead of the macroscopic optical tissue characteristics. METHODS: As existing artificial tissue phantoms designed for image forming systems are primarily targeted at wide field applications, they are unsuited for reaching the formulated objective. Therefore, a microscopy- and microendoscopy-suited artificial tissue phantom was developed and characterized. It is based on a microstructured glass surface coated with fluorescent beads at known depths covered by a scattering agent with modifiable optical properties. The phantom was examined with different kinds of microscopy systems in order to characterize its quality and stability and to demonstrate its usefulness for instrument comparison, for example, regarding structural as well as fluorescence lifetime analysis. RESULTS: The analysis of the manufactured microstructured glass surfaces showed high regularity in their physical dimensions in accordance with the specifications. Measurements of the optical parameters of the scattering medium were consistent with simulations. The fluorescent beads coating proved to be stable for a respectable period of time (about a week). The developed artificial tissue phantom was successfully used to detect differences in image quality between a research microscope and an endoscopy based system. Plausible causes for the observed differences could be derived based on the well known microstructure of the phantom. CONCLUSIONS: The artificial tissue phantom is well suited for the intended use with microscopic and microendoscopic systems. Due to its configurable design, it can be adapted to a wide range of applications. It is especially targeted at the characterization and calibration of clinical imaging systems that often lack extensive positioning capabilities such as an intrinsic z-stage.

Evaluation of hyperspectral imaging to quantify perfusion changes during the modified Allen test.

OBJECTIVES: To evaluate the capability of hyperspectral imaging (HSI), a contact-less and noninvasive technology, to monitor perfusion changes of the hand during a modified Allen test (MAT) and cuff occlusion test. Furthermore, the study aimed at obtaining objective perfusion parameters of the hand. METHODS: HSI of the hand was performed on 20 healthy volunteers with a commercially available HSI system during a MAT and a cuff occlusion test. Besides gathering red-green-blue (RGB) images, the perfusion parameters tissue hemoglobin index (THI), (superficial tissue) hemoglobin oxygenation (StO2), near-infrared perfusion (NIR), and tissue water index (TWI) were calculated for four different regions of interest on the hand. For the MAT, occlusion (OI; the ratio between the condition during occlusion and before occlusion) and reperfusion (RI; the ratio between the non-occlusion state and the prior occlusion state) indices were calculated for each perfusion parameter. All data were correlated to the clinical findings. RESULTS: False-color images showed visible differences between the various perfusion conditions during the MAT and cuff occlusion test. THI, StO2, and NIR behaved as expected from physiology, while TWI did not in the context of this study. During rest, mean THI, StO2, and NIR of the hand were 34 ± 2, 72 ± 9, and 61 ± 6, respectively. The RI for THI showed a roundabout threefold increase after reperfusion of both radial and ulnar artery and was thus, distinctly pronounced when compared with StO2 and NIR (~1.25). The OI was lowest for THI when compared with StO2 and NIR. CONCLUSIONS: HSI with its parameters THI, StO2, and NIR proved to be suitable to evaluate perfusion of the hand. By this, it could complement visual inspection during the MAT for evaluating the functionality of the superficial palmary arch before radial or ulnar artery harvest. The presented RI might deliver useful comparative values to detect pathological perfusion disorders at an early stage. As microcirculation monitoring is crucial for many medical issues, HSI shows potential to be used, besides further applications, in the monitoring of (free) flaps and transplants and microcirculation monitoring of critically ill patients.

Quenched coumarin derivatives as fluorescence lifetime phantoms for NADH and FAD.

Two-photon fluorescence lifetime imaging is a versatile laboratory technique in the field of biophotonics and its importance is also growing in the field of in vivo diagnostics for medical purposes. After years of experience in dermatology, endoscopic implementations of the technique are now posing new technical challenges. To develop, test, and compare instrumental solutions for this purpose suitable reference samples have been devised and tested. These reference samples can serve as reliable NADH- and FAD-mimicking optical phantoms for 2-photon fluorescence lifetime imaging, as they can be prepared relatively easily with reproducible and stable characteristics for this quite relevant diagnostic technique. The reference samples (mixtures of coumarin 1 and coumarin 6 in ethanol with suitable amounts of 4-hydroxy-TEMPO) have been tuned to exhibit spectral and temporal fluorescence characteristics very similar to those of NADH and FAD, the two molecules most frequently utilized to characterize cell metabolism.

Comparative investigation of reusable and single-use flexible endoscopes for urological interventions.

In order to evaluate the technical adaptability of a type of disposable endoscope compared to reusable flexible endoscopes, in vitro and in vivo studies were conducted. A disposable digital ureteroscope ("chip on tip") and two reusable endoscopes were investigated with respect to spatial resolution, geometric distortion in air and water the maximum. Additionally, the clinical performance of the disposable device was tested during clinical procedures (n = 20). The disposable endoscope showed an optical resolution of 6.72 lines/mm at 10 mm distance, similar to the other devices. In comparison, the disposable endoscope showed a barrel-shaped image distortion in air of -24.2%, which is in the middle range, but was best under water (-8.6%). The bendability of 297° (275 µm fiber) and 316° (empty channel, 1.5 F basket) and the maximum irrigation (1 m: 58.1 ml/min, 2 m: 91.9 ml/min) were convincing. Clinically the maneuverability was very good in (13/20), good or satisfactory in (7/20). Visibility was evaluated as very good in (11/20), just in (1/20) either satisfactory or sufficient. The consistency of visibility was not affected in (19/20). In all cases there were no adverse events. The technical examination and clinical application of the disposable endoscope are of equal quality compared to reusable devices. Disposable endoscopes can be an alternative to reusable devices, but economic aspects such as reduction of repair costs, sterilization effort and additional waste must be taken into account.