FluxEXPLORER: a new high-speed laser Doppler imaging system for the assessment of burn injuries.

BACKGROUND: Deep burn assessment made by clinical evaluation has an accuracy varying between 60% and 80% and will determine if a burn injury will need tangential excision and skin grafting or if it will be able to heal spontaneously. Laser Doppler Imaging (LDI) techniques allow an improved burn depth assessment but their use is limited by the time-consuming image acquisition which may take up to 6 min per image. METHODS: To evaluate the effectiveness and reliability of a newly developed full-field LDI technology, 15 consecutive patients presenting with intermediate depth burns were assessed both clinically and by FluxExplorer LDI technology. Comparison between the two methods of assessment was carried out. RESULTS: Image acquisition was done within 6 s. FluxEXPLORER LDI technology achieved a significantly improved accuracy of burn depth assessment compared to the clinical judgement performed by board certified plastic and reconstructive surgeons (P < 0.05, 93% of correctly assessed burns injuries vs. 80% for clinical assessment). CONCLUSION: Technological improvements of LDI technology leading to a decreased image acquisition time and reliable burn depth assessment allow the routine use of such devices in the acute setting of burn care without interfering with the patient's treatment. Rapid and reliable LDI technology may assist clinicians in burn depth assessment and may limit the morbidity of burn patients through a minimization of the area of surgical debridement. Future technological improvements allowing the miniaturization of the device will further ease its clinical application.

Assessment of melanocytic skin lesions with a high-definition laser Doppler imaging system.

BACKGROUND: Early detection is a major goal in the management of malignant melanoma. Besides clinical assessment many noninvasive technologies such as dermoscopy, digital dermoscopy and in vivo laser scanner microscopy are used as additional methods. Herein we tested a system to assess lesional perfusion as a tool for early melanoma detection. METHODS: Laser Doppler flow (FluxExplorer) and mole analyser (MA) score (FotoFinder) were applied to histologically verified melanocytic nevi (33) and malignant melanomas (12). RESULTS: Mean perfusion and MA scores were significantly increased in melanoma compared to nevi. However, applying an empirically determined threshold of 16% perfusion increase only 42% of the melanomas fulfilled the criterion of malignancy, whereas with the mole analyzer score 82% of the melanomas fulfilled the criterion of malignancy. CONCLUSION: Laser Doppler imaging is a highly sensitive technology to assess skin and skin tumor perfusion in vivo. Although mean perfusion is higher in melanomas compared to nevi the high numbers of false negative results hamper the use of this technology for early melanoma detection.

Application of a new laser Doppler imaging system in planning and monitoring of surgical flaps.

There is a demand for technologies able to assess the perfusion of surgical flaps quantitatively and reliably to avoid ischemic complications. The aim of this study is to test a new high-speed high-definition laser Doppler imaging (LDI) system (FluxEXPLORER, Microvascular Imaging, Lausanne, Switzerland) in terms of preoperative mapping of the vascular supply (perforator vessels) and postoperative flow monitoring. The FluxEXPLORER performs perfusion mapping of an area 9 x 9 cm with a resolution of 256 x 256 pixels within 6 s in high-definition imaging mode. The sensitivity and predictability to localize perforators is expressed by the coincidence of preoperatively assessed LDI high flow spots with intraoperatively verified perforators in nine patients. 18 free flaps are monitored before, during, and after total ischemia. 63% of all verified perforators correspond to a high flow spot, and 38% of all high flow spots correspond to a verified perforator (positive predictive value). All perfused flaps reveal a value of above 221 perfusion units (PUs), and all values obtained in the ischemic flaps are beneath 187 PU. In summary, we conclude that the present LDI system can serve as a reliable, fast, and easy-to-handle tool to detect ischemia in free flaps, whereas perforator vessels cannot be detected appropriately.

Stochastic approach to data analysis in fluorescence correlation spectroscopy.

Fluorescence correlation spectroscopy (FCS) has emerged as a powerful technique for measuring low concentrations of fluorescent molecules and their diffusion constants. In FCS, the experimental data is conventionally fit using standard local search techniques, for example, the Marquardt-Levenberg (ML) algorithm. A prerequisite for these categories of algorithms is the sound knowledge of the behavior of fit parameters and in most cases good initial guesses for accurate fitting, otherwise leading to fitting artifacts. For known fit models and with user experience about the behavior of fit parameters, these local search algorithms work extremely well. However, for heterogeneous systems or where automated data analysis is a prerequisite, there is a need to apply a procedure, which treats FCS data fitting as a black box and generates reliable fit parameters with accuracy for the chosen model in hand. We present a computational approach to analyze FCS data by means of a stochastic algorithm for global search called PGSL, an acronym for Probabilistic Global Search Lausanne. This algorithm does not require any initial guesses and does the fitting in terms of searching for solutions by global sampling. It is flexible as well as computationally faster at the same time for multiparameter evaluations. We present the performance study of PGSL for two-component with triplet fits. The statistical study and the goodness of fit criterion for PGSL are also presented. The robustness of PGSL on noisy experimental data for parameter estimation is also verified. We further extend the scope of PGSL by a hybrid analysis wherein the output of PGSL is fed as initial guesses to ML. Reliability studies show that PGSL and the hybrid combination of both perform better than ML for various thresholds of the mean-squared error (MSE).

Full-field laser Doppler perfusion imaging and monitoring with an intelligent CMOS camera.

A system for full-field laser Doppler blood flow imaging has been developed and tested on biomedical samples. The new imaging system allows 2D flow maps or monitoring flux signals to be obtained from a plurality of measured points simultaneously by using a 2D array of photodetectors. The detection part of the system is based on an intelligent CMOS camera with a built-in digital signal processor and memory. The imaging time of the system is as much as to 4 times faster than for the conventional scanning laser Doppler imager. The performance of the system was evaluated in vitro and in vivo. The first measurement results with this new system on human skin are reported.

High-speed laser Doppler perfusion imaging using an integrating CMOS image sensor.

This paper describes the design and the performance of a new high-speed laser Doppler imaging system for monitoring blood flow over an area of tissue. The new imager delivers high-resolution flow images (256x256 pixels) every 2 to 10 seconds, depending on the number of points in the acquired time-domain signal (32-512 points). This new imaging modality utilizes a digital integrating CMOS image sensor to detect Doppler signals in a plurality of points over the area illuminated by a divergent laser beam of a uniform intensity profile. The integrating property of the detector improves the signal-to-noise ratio of the measurements, which results in high-quality flow images. We made a series of measurements in vitro to test the performance of the system in terms of bandwidth, SNR, etc. Subsequently we give some examples of flow-related images measured on human skin, thus demonstrating the performance of the imager in vivo. The perspectives for future implementations of the imager for clinical and physiological applications are discussed.

Parallel single molecule detection with a fully integrated single-photon 2x2 CMOS detector array.

We present parallel single molecule detection (SMD) and fluorescence correlation spectroscopy (FCS) experiments with a fully integrated complementary metal oxide semiconductor (CMOS) single-photon 2x2 detector array. Multifocal excitation is achieved with a diffractive optical element (DOE). Special emphasis is placed on parallelization of the total system. The performance of the novel single-photon CMOS detector is investigated and compared to a state-of-the-art single-photon detecting module [having an actively quenched avalanche photodiode (APD)] by measurements on free diffusing molecules at different concentrations. Despite the order of magnitude lower detection efficiency of the CMOS detector compared to the state-of-the-art single-photon detecting module, we achieve single molecule sensitivity and reliably determine molecule concentrations. In addition, the CMOS detector performance for the determination of the fraction of slowly diffusing molecules in a primer solution (two-component analysis) is demonstrated. The potential of this new technique for high-throughput confocal-detection-based systems is discussed.