Confocal backscattering spectroscopy for leukemic and normal blood cell discrimination

Leukemia is the most common pediatric cancer and leading cause of cancer related deaths in children. Improvements in the assessment of leukemic cells have the potential to influence not only the diagnosis of leukemia, but also the risk assessment of patients during the course of the treatment, both of which are important for improving the cure rate for this disease. In this study, we report on the design and performance of a confocal laser based system built to collect backscattered light over a range of 26° at 405, 488, and 633 nm to discriminate leukemic cells from normal red blood cells (RBC) and white blood cells (WBC). The design of the system is based on the spectral differences observed from spectroscopy measurements with a similar system designed with a white light source. Significant differences are observed in the intensity and wavelength dependence of leukemic cells from normal RBC and WBC. Specifically, the distinct light scattering of RBC is due to hemoglobin absorption, allowing for its discrimination from leukemic cells, mononuclear, and polymorphonuclear WBC particularly at certain wavelengths. Meanwhile, the high scattering intensities of polymorphonuclear WBC reflect the intracellular complexity of these cells in comparison to the leukemic or normal lymphocytes. Additionally, the detected light scattering spectra for leukemic cells are consistently steeper in comparison to normal WBC, which we attributed to differences in the fractal organization of intracellular scatterers. Based on our findings, the system has potential applications in the detection and quantification of leukemic cells in blood either in vivo or in vitro, using microfluidic-based systems, for disease monitoring.