Abstract

Machine learning offers promising solutions for high-throughput single-particle analysis in label-free imaging microflow cytomtery. However, the throughput of online operations such as cell sorting is often limited by the large computational cost of the image analysis while offline operations may require the storage of an exceedingly large amount of data. Moreover, the training of machine learning systems can be easily biased by slight drifts of the measurement conditions, giving rise to a significant but difficult to detect degradation of the learned operations. We propose a simple and versatile machine learning approach to perform microparticle classification at an extremely low computational cost, showing good generalization over large variations in particle position. We present proof-of-principle classification of interference patterns projected by flowing transparent PMMA microbeads with diameters of {15.2},upmu text {m} and {18.6},upmu text {m}. To this end, a simple, cheap and compact label-free microflow cytometer is employed. We also discuss in detail the detection and prevention of machine learning bias in training and testing due to slight drifts of the measurement conditions. Moreover, we investigate the implications of modifying the projected particle pattern by means of a diffraction grating, in the context of optical extreme learning machine implementations.

Highlights

  • Machine learning offers promising solutions for high-throughput single-particle analysis in label-free imaging microflow cytomtery

  • Employing a CMOS image sensor, we acquired the interference patterns obtained by shining red laser light on transparent PMMA microparticles (with diameters of (15.2 ± 0.5)μm and (18.6 ± 0.6)μm ) flowing in a 100 μm × 100 μm microfluidic

  • It should be specified that the throughput of our setup is quite low (∼ 2.7 classified cells per second for R = 0.04 ), since our work mainly focuses on general machine learning aspects of label-free imaging flow cytometry rather than on developing a high-throughput device

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Summary

Introduction

Machine learning offers promising solutions for high-throughput single-particle analysis in label-free imaging microflow cytomtery. The moving particles are illuminated, usually by a laser, and the corresponding forward and/or side-scattering intensities are measured, together with the fluorescent emission of selectively attached probes These devices are widely used to investigate the structure and the chemical composition of large populations of cells in many applications concerning life science and clinical diagnosis. This limitation can be overcome, at the cost of increasing system and instrumentation complexity, by encoding optical spatial information into a temporal sequence that is measured by a single photodetector An application of this technique, named Serial Time-Encoded Amplified Microscopy (STEAM), combines the wide spectral bandwidth of a femtosecond pulse laser with both temporal and spatial dispersive optical elements achieving label-free single cell imaging at a very high throughput, up to ∼ 100, 000 cells/s10–12

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