RESUMEN
Due to the shortage of healthy donor organs, steatotic livers are commonly used for transplantation, placing patients at higher risk for graft dysfunction and lower survival rates. Raman Spectroscopy is a technique which has shown the ability to rapidly detect the vibration state of C-H bonds in triglycerides. The aim of this study is to determine whether conventional Raman spectroscopy can reliably detect and quantify fat in an animal model of liver steatosis. Mice and rats fed a methionine and choline-deficient (MCD) and control diets were sacrificed on one, two, three and four weeks' time points. A confocal Raman microscope, a commercial Raman (iRaman) fiber optic probe and a highly sensitive Raman fiber optic probe system, the latter utilizing a 785 nm excitation laser, were used to detect changes in the Raman spectra of steatotic mouse livers. Thin layer chromatography was used to assess the triglyceride content of liver specimens, and sections were scored blindly for fat content using histological examination. Principal component analysis (PCA) of Raman spectra was used to extract the principal components responsible for spectroscopic differences with MCD week (time on MCD diet). Confocal Raman microscopy revealed the presence of saturated fats in mice liver sections. A commercially available handheld Raman spectroscopy probe could not distinguish the presence of fat in the liver whereas our specially designed, high throughput Raman system could clearly distinguish lobe-specific changes in fat content. In the left lobe in particular, the Raman PC scores exhibited a significant correlation (R(2) = 0.96) with the gold standard, blinded scoring by histological examination. The specially designed, high throughput Raman system can be used for clinical purposes. Its application to the field of transplantation would enable surgeons to determine the hepatic fat content of the donor's liver in the field prior to proceeding with organ retrieval. Next steps include validating these results in a prospective analysis of human liver transplantation implant biopsies.
Asunto(s)
Hígado Graso/diagnóstico , Tecnología de Fibra Óptica , Espectrometría Raman/instrumentación , Animales , Modelos Animales de Enfermedad , Endoscopía , Hígado Graso/metabolismo , Hígado Graso/patología , Humanos , Masculino , Ratones , RatasRESUMEN
The management of cancer in the periphery lung is in critical need of new strategies. Here, the development and test of a novel miniature Raman probe capable of navigating the peripheral lung architecture is reported. The probe was 1.35 mm in diameter, with a minimum bend radius of 13 mm and had a large light collection area for its size. Peripheral lung Raman spectra were successfully obtained from normal tissue and cancerous nodule using the probe coupled to a home-made rapid Raman spectroscopy system with a fast integration time of 1 second and a low excitation power of 15 mW. This is the first time in vivo Raman spectra from the periphery lung being reported. The collected spectra showed lipid, protein and deoxyhemoglobin signatures that might be useful for classifying pathology. Large scale clinical study is planned to confirm the utility of this new technology for improving periphery lung cancer detection. Left: Radial ultrasound image of a peripheral lung nodule: size given by crosshairs D1 and D2. Right: Truncated Raman spectra of a cancerous nodule, whole blood, and normal peripheral airway tissue. Spectra were shifted on intensity scale for clarity.
Asunto(s)
Detección Precoz del Cáncer/instrumentación , Neoplasias Pulmonares/diagnóstico , Miniaturización/instrumentación , Espectrometría Raman/instrumentación , Diseño de Equipo , HumanosRESUMEN
Currently the most sensitive method for localizing lung cancers in central airways is autofluorescence bronchoscopy (AFB) in combination with white light bronchoscopy (WLB). The diagnostic accuracy of WLB + AFB for high grade dysplasia (HGD) and carcinoma in situ is variable depending on physician's experience. When WLB + AFB are operated at high diagnostic sensitivity, the associated diagnostic specificity is low. Raman spectroscopy probes molecular vibrations and gives highly specific, fingerprint-like spectral features and has high accuracy for tissue pathology classification. In this study we present the use of a real-time endoscopy Raman spectroscopy system to improve the specificity. A spectrum is acquired within 1 second and clinical data are obtained from 280 tissue sites (72 HGDs/malignant lesions, 208 benign lesions/normal sites) in 80 patients. Using multivariate analyses and waveband selection methods on the Raman spectra, we have demonstrated that HGD and malignant lung lesions can be detected with high sensitivity (90%) and good specificity (65%).