Raman spectroscopy is well suited for readily revealing information about bio-samples. As such, this technique has been applied to a wide range of areas, particularly in bio-medical diagnostics. Raman scattering in bio-samples typically has a low signal level due to the nature of inelastic scattering of photons. To achieve a high signal level, usually a high numerical aperture objective is employed. One drawback with these objectives is that their working distance is very short. However, in many cases of clinical diagnostics, a long working distance is preferable. We propose a practical solution to this problem by enhancing the Raman signal using a parabolic reflector. The high signal level is achieved through the large light collection solid angle of the parabolic reflector while the long working distance is ensured by the novel design of our microscope. The enhancement capability of the microscope was demonstrated on four types of samples. Among these samples, we find that this microscope design is most suitable for turbid samples.
Raman spectroscopy has demonstrated great potential for skin wound assessment. Given that biochemical changes in wound healing is depth dependent as the skin is a layered structure, depth sensitive Raman spectroscopy could enhance the power of Raman spectroscopy in this application. Considering the critical importance of rodent studies in the field of skin wound assessment, it is necessary to develop and validate a system that can perform depth sensitive measurements in rat skin with a proper target depth range. In this manuscript, we report the design, optimization and evaluation of a new snapshot depth-sensitive Raman instrument for rat skin measurements. The optical design and optimization process are presented first. The depth sensitive measurement performance is characterized on both ex vivo porcine skin with a gradient of layer thickness and ex vivo rat skin samples with wounds. The statistical analysis of the measured Raman spectra demonstrates the feasibility of differentiation between the wound edge and healthy skin. Moreover, the accuracy of classification improves monotonically as more data from new depths are used, which implies that each depth offers additional information useful for classification. This instrument demonstrates the ability to perform snapshot depth sensitive Raman measurements from rat skin, which paves the way towards in vivo preclinical studies of rat skin wounds.
Raman spectroscopy has demonstrated its great potential in skin wound assessment. Given that biochemical changes in skin wound healing is a layer dependent process, depth sensitive Raman spectroscopy could enhance the power of Raman spectroscopy in this application. Considering the critical importance of rodent studies in the field of skin wound assessment, it is necessary to develop and validate a system that can perform depth sensitive measurements in rat skin with a proper target depth range. In this manuscript, we report the design, optimization and evaluation of a new snapshot depth-sensitive Raman instrument for rat skin measurements. The optical design and optimization process are presented first. The depth sensitive measurement performance is characterized on ex vivo rat skin samples with wounds. Raman signal emitted by the ex vivo rat skin from different target depths were simultaneously acquired. The feasibility of using the measured Raman spectra to differentiate between the wound edge and healthy skin was validated using PLS-LDA with leave-one-out. The accuracy of the classification improves monotonically as more data from new depths are used, which implies that each depth offers additional information useful for classification. This instrument demonstrates the ability to perform snapshot depth sensitive Raman measurements from rat skin, which paves the way towards in vivo preclinical studies of rat skin wounds.
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