Fluorescence photobleaching of urine for improved signal to noise ratio of the Raman signal – An exploratory study

Abstract

Urine analysis is an important clinical test routinely performed in pathology labs for disease diagnosis and prognosis. In recent years, near-infrared Raman spectroscopy has drawn considerable attention for urine analysis as it can provide rapid, reliable, and reagent-free analysis of urine samples. However, one important practical problem encountered in such Raman measurements is the orders of magnitude stronger spectral background preventing one to utilize the full dynamic range of the detector which is required for the measurement of Raman signal with good signal-to-noise ratio (SNR). We report here the results of an exploratory study carried out on human urine samples to show that the photobleaching, which is a major disadvantage during the fluorescence measurement, could be utilized for suppressing the measured background to improve the SNR of the Raman peaks. It was found that once the photobleaching reached its plateau, there were improvements by ~67% and ~47% in the SNR and the signal to background ratio (SBR), respectively, of the Raman signals as compared to the spectra measured at the start of acquisition. Further, the reduced background also allowed us to utilize the full dynamic range of the detector at increased integration time without saturating the detector indicating the possibility of obtaining an improved detection limit.