Digital holographic microscopy for detection of Trypanosoma cruzi parasites in fresh blood mounts

Abstract

An off-axis holographic microscope, in a transmission mode, calibrated to automatically detect the presence of Trypanosoma cruzi in blood is developed as an alternative diagnosis tool for Chagas disease. Movements of the microorganisms are detected by measuring the phase shift they produce on the transmitted wave front. A thin layer of blood infected by Trypanosoma cruzi parasites is examined in the holographic microscope, the images of the visual field being registered with a CCD camera. Two consecutive holograms of the same visual field are subtracted point by point and a phase contrast image of the resulting hologram is reconstructed by means of the angular spectrum propagation algorithm. This method enables the measurement of phase distributions corresponding to temporal differences between digital holograms in order to detect whether parasites are present or not. Experimental results obtained using this technique show that it is an efficient alternative that can be incorporated successfully as a part of a fully automatic system for detection and counting of this type of microorganisms.