Multispectral imaging of healthy and diseased red blood cells using confocal microscopy

Abstract

Red blood cell disorders represent the most common single-gene defects and pose a major public health problem, particularly in tropical countries, occurring with high frequency. Their diagnose can sometimes be difficult due to the coexistence of different causes of anemia, such as thalassemia and iron deficiency, and blood transfusions, among other factors, and requires expensive and complex molecular tests. In this work, blood samples from patients with different syndromes of alphathalassemia and iron deficiency (including anemia) as well as healthy (control) subjects were analyzed under a Leica TCS SP8 confocal microscope Samples exhibited autofluorescence when excited at 405 nm and three experimental descriptors calculated from the mean emission intensities at 502 nm, 579 nm, 628 nm, and 649 nm allowed us to discriminate between diseased and healthy cells. According to the results obtained, spectral confocal microscopy could serve as a tool in the diagnosis of thalassemia.