Fast and robust Fourier domain-based classification for on-chip lens-free flow cytometry.

Abstract

The development of portable haematology analysers receives increased attention due to their deployability in resource-limited or emergency settings. Lens-free in-line holographic microscopy is one of the technologies that is being pushed forward in this regard as it eliminates complex and expensive optics, making miniaturisation and integration with microfluidics possible. On-chip flow cytometry enables high-speed capturing of individual cells in suspension, giving rise to high-throughput cell counting and classification. To perform a real-time analysis on this high-throughput content, we propose a fast and robust framework for the classification of leukocytes. The raw data consists of holographic acquisitions of leukocytes, captured with a high-speed camera as they are flowing through a microfluidic chip. Three different types of leukocytes are considered: granulocytes, monocytes and T-lymphocytes. The proposed method bypasses the reconstruction of the holographic data altogether by extracting Zernike moments directly from the frequency domain. By doing so, we introduce robustness to translations and rotations of cells, as well as to changes in distance of a cell with respect to the image sensor, achieving classification accuracies up to 96.8%. Furthermore, the reduced computational complexity of this approach, compared to traditional frameworks that involve the reconstruction of the holographic data, allows for very fast processing and classification, making it applicable in high-throughput flow cytometry setups.