Hyperspectral imaging has been explored for clinical applications in various medical disciplines. Based on our experiences, the potential of this versatile imaging method as well as pitfalls and limitations of current approaches are discussed. The use of reference samples and simple image modeling strategies are suggested to avoid misinterpretation and achieve a more conclusive image interpretation.
Hyperspectral imaging (HSI) is a new emerging modality for noncontact assessement of tissue perfusion parameters. This study includes investigations on monitoring perfusion changes of the hand during a modified Allen test and cuff occlusion test, on the influence of the skin tones during HSI-measurements and the feasibility of HSI monitoring of intraoral free flaps
The utilization of lasers in dentistry expands greatly in recent years. For instance, fs-lasers are effective for both drilling and caries prevention, while cw-lasers are useful for adhesive hardening. A cutting-edge application of lasers in dentistry is the debonding of veneers. While there are pre-existing tools for this purpose, there is still potential for improvement. Initial efforts to investigate laser assisted debonding mechanisms with measurements of the optical and mechanical properties of teeth and prosthetic ceramics are presented. Preliminary tests conducted with a laser system used for debonding that is commercially available showed differences in the output power set at the systems console to that at specified distances from the handpiece. Furthermore, the optical properties of the samples (human teeth and ceramics) were characterised. The optical properties of the ceramics should closely resemble those of teeth in terms of look and feel, but they also influence the laser assisted debonding technique and thus must be taken into account. In addition first attempts were performed to investigate the mechanical properties of the samples by means of pump-probe-elastography under a microscope. By analyzing the sample surface up to 20 ns after a fs-laser pulse impact, pressure and shock waves could be detected, which can be utilized to determine the elastic constants of specific materials. Together such investigations are needed to shape the basis for a purely optical approach of debonding of veneers utilizing acoustic waves.
The perfusion in cutaneous flap transplants needs to be monitored in order to detect vascular problems as early as possible. This can avoid tissue hypoxia and therefore, necrosis of the transplant. Since free flap transplant failures tend to happen more commonly during surgery than at a later onset, a non-contact real-time imaging device would be most advantageous. As hyperspectral imaging (HSI) is a new emerging modality to asses free flap perfusion contactless, this study aimed to investigate whether perfusion data can be interpreted appropriately using HSI, especially in regard to the individual skin tone. Further factors that might alter these HSI-interpretations, such as aging, BMI, different sexes or smoking habits, were also considered. Therefore, a prospective feasibility study was conducted, including 101 volunteers from whom images were taken on 16 different body sites. Skin pigmentation classification was performed using the Fitzpatrick skin type classification questionnaire and the individual typology angle (ITA) acquired from the images. Perfusion indices provided by the camera software were correlated to the possible influencing factors. The results show that a dark skin tone related to a high amount of melanin may influence the HSI-measurements and thus changes the HSI-derived perfusion indices. In addition, certain physiological influencing factors such as age, BMI and sex alter the tissue composition and qualities, thus showing measurement peculiarities within these groups. In conclusion, hyperspectral imaging can be used for perfusion assessment for people with lighter skin tone levels. Further developments are appreciated especially regarding skin pigmentation and the interpretation of indices of greater skin tone levels.
Existing optical tissue phantoms are usually designed for wide field imaging systems and not readily usable for microscopic or endoscopic systems, especially such without any z-stage. Therefore a fs-laser microstructured artificial tissue phantom with adaptable geometric, tissue-optical and localized fluorescence properties enabling comparison and testing of different microscopic/endoscopic systems was designed, characterized and tested.
FLIM of metabolic coenzymes, as NAD(P)H and FAD, is now widely accepted to be one of the most important imaging methods for cell metabolism. However, new algorithms are needed to circumvent various problems and to image cell metabolism and redox state from fluorescence lifetimes. The significance of a metabolic index based on NAD(P)H FLIM will be explained and compared with the fluorescence lifetime induced redox ratio (FLIRR). The importance of FMN will be discussed and the FLIRR approach will be extended. Using a three channel TCSPC system simultaneous metabolic NADH/FAD/FMN FLIM and oxygen PLIM/dFLIM (delayed fluorescence) could be realized.
Liquid reference samples for 2-photon fluorescence lifetime investigations were developed, based on coumarin 1 and 6 and 4-hydroxy-TEMPO as quencher. Spectral and fluorescence lifetime characteristics similar to intracellular NADH and FAD were achieved.
Hyperspectral imaging to monitor perfusion parameters during the Allen test was performed in 20 volunteers. Clear differences in tissue oxygenation and hemoglobin index during rest, occlusion and reperfusion were visually and computationally evident.
Thermography is used in many application areas like non-destructive testing, architecture or zoology. Approaches to use thermography for medical diagnostics are usually focused on the detection of tumors. The usefulness and specificity for this purpose has been debated. Nevertheless it seems reasonable to evaluate the applicability of thermography as a tool for determining temperature distributions on the surface of, e.g., light diffusers or biological tissue in the field of laser medicine. The assessment of tissue-heating in treatments like interstitial photodynamic therapy (iPDT) is of particular interest since additional tissue damage due to tissue heating is usually undesired and has thus to be avoided. Monte Carlo based simulations of the expected heat distribution in tissue are based on idealized setups. They usually omit certain characteristics of fibers, like inhomogeneities in a light diffuser or losses at components such as internal mirrors, that might potentially result in local hot spots and therefore in heating beyond an acceptable temperature. Experimental techniques based on thermocouples, or similar point-based temperature measurement devices, provide only a limited view of the temperature distribution inside or at the surface of the treated tissue. As a high resolution non-contact wide-field technique thermography although has some limitations. We present an evaluation of the usefulness of thermographic methods for recording the heat distribution on tissue and tissue phantom surfaces during laser treatment in an iPDT-like setup and compare the results with Monte Carlo based simulations.
Interstitial photodynamic therapy (iPDT) is currently being investigated as a light-based treatment option for highly malignant brain tumours (glioblastomas/GBM). To obtain a sufficient irradiation of the tumour, quantitative knowledge about the light propagation in the tissue is required for the light dosimetry calculations underlying the clinical treatment planning. To individualize the light dosimetry calculations, the optical properties of the irradiated tissue need to be determined in-vivo. A novel approach for this purpose is based on the direction-resolved light detection within the tissue, using a rotating optical side-view probe. During measurement, the tissue is irradiated via a separate interstitially placed light applicator, and from the angular dependence of the recorded signal the optical tissue properties are calculated, based on a solution of the radiative transfer equation (RTE). Measurements were performed on liquid tissue phantoms and biological tissue samples. As a result, an over- and underestimation of the calculated optical absorption and scattering coefficients may arise in some situations, but the effective attenuation coefficient remains largely unaffected and corresponds well with literature values.
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