Abstract
.We report the ex vivo results of an in-house fabricated portable device based on polarized fluorescence measurements in the clinical environment. This device measures the polarized fluorescence and elastic scattering spectra with 405-nm laser and white light sources, respectively. The dominating fluorophore with 405-nm excitation is flavin adenine dinucleotide (FAD) with a fluorescence peak around 510 nm. The measured spectra are highly modulated by the interplay of scattering and absorption effects. Due to this, valuable information gets masked. To reduce these effects, intrinsic fluorescence was extracted by normalizing polarized fluorescence spectra with polarized elastic scattering spectra obtained. A number of fluorophores contribute to the fluorescence spectra and need to be decoupled to understand their roles in the progression of cancer. Nelder–Mead method has been utilized to fit the spectral profile with Gaussian to decouple the different bands of contributing fluorophores (FAD and porphyrin). The change in concentration of FAD during disease progression manifests in the change in ratio of total area to FWHM of its Gaussian profile. Receiver operating characteristic (ROC) curve analysis has been used to discriminate different grades of cervical precancer by using the ratio as input parameter. The sensitivity and specificity for discrimination of normal samples from CIN I (cervical intraepithelial neoplasia) are 75% and 54%, respectively. Further, the normal samples can be discriminated from CIN II samples with 100% and 82% sensitivity and specificity, respectively, and the CIN I from CIN II samples can also be discriminated with 100% sensitivity and 90% specificity, respectively. The results show that the change in the concentration of (FAD) can be used as a marker to discriminate the different grades of the cancer and biochemical changes at an early stage of the cancer can also be monitored with this technique.
Highlights
Fluorescence spectroscopy is a sensitive technique, applied extensively for detection of different types of cancers
Several groups have worked on diagnosis of cervical precancer using laser-induced fluorescence spectroscopy[14,15] and in combination with reflectance spectroscopy[16,17,18] for comparison and to improve the detection efficacy
Meena et al.: Concentration of flavin adenine dinucleotide (FAD) as a marker for cervical precancer detection fluorophores contribute to the broad fluorescence spectrum when tissue is excited at 405 nm
Summary
Fluorescence spectroscopy is a sensitive technique, applied extensively for detection of different types of cancers. Meena et al.: Concentration of FAD as a marker for cervical precancer detection fluorophores contribute to the broad fluorescence spectrum when tissue is excited at 405 nm. These fluorophores need to be decoupled to understand their roles in the progression of cancer.[35] Nelder–Mead method has been utilized to fit the spectral profile with Gaussian to decouple the different bands of contributing fluorophores (FAD and porphyrin).[20,31] The change in concentration of FAD during disease progression manifests in the change in ratio of total area and FWHM of its Gaussian profile. Receiver operating characteristic (ROC) curve analysis has been used to discriminate different grades of cervical precancer by using the ratio as input parameter
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