Detection of acute promyelocytic leukemia in peripheral blood and bone marrow with annotation-free deep learning.

While optical microscopy inspection of blood films and bone marrow aspirates by a hematologist is a crucial step in establishing diagnosis of acute leukemia, especially in low-resource settings where other diagnostic modalities are not available, the task remains time-consuming and prone to human inconsistencies. This has an impact especially in cases of Acute Promyelocytic Leukemia (APL) that require urgent treatment. Integration of automated computational hematopathology into clinical workflows can improve the throughput of these services and reduce cognitive human error. However, a major bottleneck in deploying such systems is a lack of sufficient cell morphological object-labels annotations to train deep learning models. We overcome this by leveraging patient diagnostic labels to train weakly-supervised models that detect different types of acute leukemia. We introduce a deep learning approach, Multiple Instance Learning for Leukocyte Identification (MILLIE), able to perform automated reliable analysis of blood films with minimal supervision. Without being trained to classify individual cells, MILLIE differentiates between acute lymphoblastic and myeloblastic leukemia in blood films. More importantly, MILLIE detects APL in blood films (AUC 0.94 ± 0.04) and in bone marrow aspirates (AUC 0.99 ± 0.01). MILLIE is a viable solution to augment the throughput of clinical pathways that require assessment of blood film microscopy.

Content aware multi-focus image fusion for high-magnification blood film microscopy.

Automated digital high-magnification optical microscopy is key to accelerating biology research and improving pathology clinical pathways. High magnification objectives with large numerical apertures are usually preferred to resolve the fine structural details of biological samples, but they have a very limited depth-of-field. Depending on the thickness of the sample, analysis of specimens typically requires the acquisition of multiple images at different focal planes for each field-of-view, followed by the fusion of these planes into an extended depth-of-field image. This translates into low scanning speeds, increased storage space, and processing time not suitable for high-throughput clinical use. We introduce a novel content-aware multi-focus image fusion approach based on deep learning which extends the depth-of-field of high magnification objectives effectively. We demonstrate the method with three examples, showing that highly accurate, detailed, extended depth of field images can be obtained at a lower axial sampling rate, using 2-fold fewer focal planes than normally required.

Optical mesoscopy, machine learning, and computational microscopy enable high information content diagnostic imaging of blood films.

Automated image-based assessment of blood films has tremendous potential to support clinical haematology within overstretched healthcare systems. To achieve this, efficient and reliable digital capture of the rich diagnostic information contained within a blood film is a critical first step. However, this is often challenging, and in many cases entirely unfeasible, with the microscopes typically used in haematology due to the fundamental trade-off between magnification and spatial resolution. To address this, we investigated three state-of-the-art approaches to microscopic imaging of blood films which leverage recent advances in optical and computational imaging and analysis to increase the information capture capacity of the optical microscope: optical mesoscopy, which uses a giant microscope objective (Mesolens) to enable high-resolution imaging at low magnification; Fourier ptychographic microscopy, a computational imaging method which relies on oblique illumination with a series of LEDs to capture high-resolution information; and deep neural networks which can be trained to increase the quality of low magnification, low resolution images. We compare and contrast the performance of these techniques for blood film imaging for the exemplar case of Giemsa-stained peripheral blood smears. Using computational image analysis and shape-based object classification, we demonstrate their use for automated analysis of red blood cell morphology and visualization and detection of small blood-borne parasites such as the malarial parasite Plasmodium falciparum. Our results demonstrate that these new methods greatly increase the information capturing capacity of the light microscope, with transformative potential for haematology and more generally across digital pathology. © 2021 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.

Wide-field parallel mapping of local spectral and topographic information with white light interference microscopy.

Fourier analysis of interferograms captured in white light interference microscopy is proposed for performing simultaneous local spectral and topographic measurements at high spatial resolution over a large field of view. The technique provides a wealth of key information on local sample properties. We describe the processing and calibration steps involved to produce reflectivity maps of spatially extended samples. This enables precise and fast identification between different materials at a local scale of 1 µm. We also show that the recovered spectral information can be further used for improving topography measurements, particularly in the case of samples combining dielectric and conducting materials in which the complex refractive index can result in nanometric height errors.

Structure-dependent amplification for denoising and background correction in Fourier ptychographic microscopy.

Fourier Ptychographic Microscopy (FPM) allows high resolution imaging using iterative phase retrieval to recover an estimate of the complex object from a series of images captured under oblique illumination. FPM is particularly sensitive to noise and uncorrected background signals as it relies on combining information from brightfield and noisy darkfield (DF) images. In this article we consider the impact of different noise sources in FPM and show that inadequate removal of the DF background signal and associated noise are the predominant cause of artefacts in reconstructed images. We propose a simple solution to FPM background correction and denoising that outperforms existing methods in terms of image quality, speed and simplicity, whilst maintaining high spatial resolution and sharpness of the reconstructed image. Our method takes advantage of the data redundancy in real space within the acquired dataset to boost the signal-to-background ratio in the captured DF images, before optimally suppressing background signal. By incorporating differentially denoised images within the classic FPM iterative phase retrieval algorithm, we show that it is possible to achieve efficient removal of background artefacts without suppression of high frequency information. The method is tested using simulated data and experimental images of thin blood films, bone marrow and liver tissue sections. Our approach is non-parametric, requires no prior knowledge of the noise distribution and can be directly applied to other hardware platforms and reconstruction algorithms making it widely applicable in FPM.

Data-driven malaria prevalence prediction in large densely populated urban holoendemic sub-Saharan West Africa.

Over 200 million malaria cases globally lead to half-million deaths annually. The development of malaria prevalence prediction systems to support malaria care pathways has been hindered by lack of data, a tendency towards universal "monolithic" models (one-size-fits-all-regions) and a focus on long lead time predictions. Current systems do not provide short-term local predictions at an accuracy suitable for deployment in clinical practice. Here we show a data-driven approach that reliably produces one-month-ahead prevalence prediction within a densely populated all-year-round malaria metropolis of over 3.5 million inhabitants situated in Nigeria which has one of the largest global burdens of P. falciparum malaria. We estimate one-month-ahead prevalence in a unique 22-years prospective regional dataset of > 9 × 104 participants attending our healthcare services. Our system agrees with both magnitude and direction of the prediction on validation data achieving MAE ≤ 6 × 10-2, MSE ≤ 7 × 10-3, PCC (median 0.63, IQR 0.3) and with more than 80% of estimates within a (+ 0.1 to - 0.05) error-tolerance range which is clinically relevant for decision-support in our holoendemic setting. Our data-driven approach could facilitate healthcare systems to harness their own data to support local malaria care pathways.

Expert-level automated malaria diagnosis on routine blood films with deep neural networks.

Over 200 million malaria cases globally lead to half a million deaths annually. Accurate malaria diagnosis remains a challenge. Automated imaging processing approaches to analyze Thick Blood Films (TBF) could provide scalable solutions, for urban healthcare providers in the holoendemic malaria sub-Saharan region. Although several approaches have been attempted to identify malaria parasites in TBF, none have achieved negative and positive predictive performance suitable for clinical use in the west sub-Saharan region. While malaria parasite object detection remains an intermediary step in achieving automatic patient diagnosis, training state-of-the-art deep-learning object detectors requires the human-expert labor-intensive process of labeling a large dataset of digitized TBF. To overcome these challenges and to achieve a clinically usable system, we show a novel approach. It leverages routine clinical-microscopy labels from our quality-controlled malaria clinics, to train a Deep Malaria Convolutional Neural Network classifier (DeepMCNN) for automated malaria diagnosis. Our system also provides total Malaria Parasite (MP) and White Blood Cell (WBC) counts allowing parasitemia estimation in MP/µL, as recommended by the WHO. Prospective validation of the DeepMCNN achieves sensitivity/specificity of 0.92/0.90 against expert-level malaria diagnosis. Our approach PPV/NPV performance is of 0.92/0.90, which is clinically usable in our holoendemic settings in the densely populated metropolis of Ibadan. It is located within the most populous African country (Nigeria) and with one of the largest burdens of Plasmodium falciparum malaria. Our openly available method is of importance for strategies aimed to scale malaria diagnosis in urban regions where daily assessment of thousands of specimens is required.

Digital refocusing and extended depth of field reconstruction in Fourier ptychographic microscopy.

Fourier ptychography microscopy (FPM) is a recently developed microscopic imaging method that allows the recovery of a high-resolution complex image by combining a sequence of bright and darkfield images acquired under inclined illumination. The capacity of FPM for high resolution imaging at low magnification makes it particularly attractive for applications in digital pathology which require imaging of large specimens such as tissue sections and blood films. To date most applications of FPM have been limited to imaging thin samples, simplifying both image reconstruction and analysis. In this work we show that, for samples of intermediate thickness (defined here as less than the depth of field of a raw captured image), numerical propagation of the reconstructed complex field allows effective digital refocusing of FPM images. The results are validated by comparison against images obtained with an equivalent high numerical aperture objective lens. We find that post reconstruction refocusing (PRR) yields images comparable in quality to adding a defocus term to the pupil function within the reconstruction algorithm, while reducing computing time by several orders of magnitude. We apply PRR to visualize FPM images of Giemsa-stained peripheral blood films and present a novel image processing pipeline to construct an effective extended depth of field image which optimally displays the 3D sample structure in a 2D image. We also show how digital refocusing allows effective correction of the chromatic focus shifts inherent to the low magnification objective lenses used in FPM setups, improving the overall quality of color FPM images.

Coherence scanning interferometry allows accurate characterization of micrometric spherical particles contained in complex media.

Characterizing very small particles, from a few dozen micrometers to the nanometric scale, is a very challenging application in a wide range of domains. In this work, we demonstrate, through the recovery of silica and polystyrene bead properties (i.e. their size and refractive index) that Coherence Scanning Interferometry (CSI), in addition of being contactless, non-destructive, label-free and very well spatially resolved, is a very interesting and promising tool for such complex characterization. The CSI system is used as an imaging Fourier transform spectrometer meaning that the characterizations are achieved by analyzing the interference signal in the spectral domain. Some simulations of the proposed technique are presented and show that the accuracy of such characterization, in particular the measurement of the refractive index, are closely related to the signal to noise ratio. This observation is thereafter confirmed by the experimental results of beads buried within the depth of a transparent sample. Finally, the method is theoretically tested in the case of a scattering medium in which the quality of the signal is highly degraded. In this context, a geometrical approach enabling the simulation of an interference signal from a scattering layer is first proposed and then validated by means of comparison with experimental data.

Depth-resolved local reflectance spectra measurements in full-field optical coherence tomography.

Full-field optical coherence tomography (FF-OCT) is a widely used technique for applications such as biological imaging, optical metrology, and materials characterization, providing structural and spectral information. By spectral analysis of the backscattered light, the technique of spectroscopic-OCT enables the differentiation of structures having different spectral properties, but not the determination of their reflectance spectrum. For surface measurements, this can be achieved by applying a Fourier transform to the interferometric signals and using an accurate calibration of the optical system. An extension of this method is reported for local spectroscopic characterization of transparent samples and in particular for the determination of depth-resolved reflectance spectra of buried interfaces. The correct functioning of the method is demonstrated by comparing the results with those obtained using a program based on electromagnetic matrix methods for stratified media. Experimental measurements of spatial resolutions are provided to demonstrate the smallest structures that can be characterized.