Kanghyun Kim

Lucas Kreiss, Shaowei Jiang, Xiang Li et al. · pku

Until recently, conventional biochemical staining had the undisputed status as well-established benchmark for most biomedical problems related to clinical diagnostics, fundamental research and biotechnology. Despite this role as gold-standard, staining protocols face several challenges, such as a need for extensive, manual processing of samples, substantial time delays, altered tissue homeostasis, limited choice of contrast agents for a given sample, 2D imaging instead of 3D tomography and many more. Label-free optical technologies, on the other hand, do not rely on exogenous and artificial markers, by exploiting intrinsic optical contrast mechanisms, where the specificity is typically less obvious to the human observer. Over the past few years, digital staining has emerged as a promising concept to use modern deep learning for the translation from optical contrast to established biochemical contrast of actual stainings. In this review article, we provide an in-depth analysis of the current state-of-the-art in this field, suggest methods of good practice, identify pitfalls and challenges and postulate promising advances towards potential future implementations and applications.

Shiqi Xu, Wenhui Liu, Xi Yang et al.

Fast noninvasive probing of spatially varying decorrelating events, such as cerebral blood flow beneath the human skull, is an essential task in various scientific and clinical settings. One of the primary optical techniques used is diffuse correlation spectroscopy (DCS), whose classical implementation uses a single or few single-photon detectors, resulting in poor spatial localization accuracy and relatively low temporal resolution. Here, we propose a technique termed Classifying Rapid decorrelation Events via Parallelized single photon dEtection (CREPE)}, a new form of DCS that can probe and classify different decorrelating movements hidden underneath turbid volume with high sensitivity using parallelized speckle detection from a $32\times32$ pixel SPAD array. We evaluate our setup by classifying different spatiotemporal-decorrelating patterns hidden beneath a 5mm tissue-like phantom made with rapidly decorrelating dynamic scattering media. Twelve multi-mode fibers are used to collect scattered light from different positions on the surface of the tissue phantom. To validate our setup, we generate perturbed decorrelation patterns by both a digital micromirror device (DMD) modulated at multi-kilo-hertz rates, as well as a vessel phantom containing flowing fluid. Along with a deep contrastive learning algorithm that outperforms classic unsupervised learning methods, we demonstrate our approach can accurately detect and classify different transient decorrelation events (happening in 0.1-0.4s) underneath turbid scattering media, without any data labeling. This has the potential to be applied to noninvasively monitor deep tissue motion patterns, for example identifying normal or abnormal cerebral blood flow events, at multi-Hertz rates within a compact and static detection probe.

Shiqi Xu, Xiang Dai, Xi Yang et al.

We report Tensorial Tomographic Differential Phase-Contrast microscopy (T2DPC), a quantitative label-free tomographic imaging method for simultaneous measurement of phase and anisotropy. T2DPC extends differential phase-contrast microscopy, a quantitative phase imaging technique, to highlight the vectorial nature of light. The method solves for permittivity tensor of anisotropic samples from intensity measurements acquired with a standard microscope equipped with an LED matrix, a circular polarizer, and a polarization-sensitive camera. We demonstrate accurate volumetric reconstructions of refractive index, birefringence, and orientation for various validation samples, and show that the reconstructed polarization structures of a biological specimen are predictive of pathology.

Xing Yao, Vinayak Pathak, Haoran Xi et al.

This work demonstrates a multi-lens microscopic imaging system that overlaps multiple independent fields of view on a single sensor for high-efficiency automated specimen analysis. Automatic detection, classification and counting of various morphological features of interest is now a crucial component of both biomedical research and disease diagnosis. While convolutional neural networks (CNNs) have dramatically improved the accuracy of counting cells and sub-cellular features from acquired digital image data, the overall throughput is still typically hindered by the limited space-bandwidth product (SBP) of conventional microscopes. Here, we show both in simulation and experiment that overlapped imaging and co-designed analysis software can achieve accurate detection of diagnostically-relevant features for several applications, including counting of white blood cells and the malaria parasite, leading to multi-fold increase in detection and processing throughput with minimal reduction in accuracy.

Kanghyun Kim, Pavan Chandra Konda, Colin L. Cooke et al.

Standard microscopes offer a variety of settings to help improve the visibility of different specimens to the end microscope user. Increasingly, however, digital microscopes are used to capture images for automated interpretation by computer algorithms (e.g., for feature classification, detection or segmentation), often without any human involvement. In this work, we investigate an approach to jointly optimize multiple microscope settings, together with a classification network, for improved performance with such automated tasks. We explore the interplay between optimization of programmable illumination and pupil transmission, using experimentally imaged blood smears for automated malaria parasite detection, to show that multi-element "learned sensing" outperforms its single-element counterpart. While not necessarily ideal for human interpretation, the network's resulting low-resolution microscope images (20X-comparable) offer a machine learning network sufficient contrast to match the classification performance of corresponding high-resolution imagery (100X-comparable), pointing a path towards accurate automation over large fields-of-view.

Colin L. Cooke, Fanjie Kong, Amey Chaware et al.

This paper introduces a new method of data-driven microscope design for virtual fluorescence microscopy. Our results show that by including a model of illumination within the first layers of a deep convolutional neural network, it is possible to learn task-specific LED patterns that substantially improve the ability to infer fluorescence image information from unstained transmission microscopy images. We validated our method on two different experimental setups, with different magnifications and different sample types, to show a consistent improvement in performance as compared to conventional illumination methods. Additionally, to understand the importance of learned illumination on inference task, we varied the dynamic range of the fluorescent image targets (from one to seven bits), and showed that the margin of improvement for learned patterns increased with the information content of the target. This work demonstrates the power of programmable optical elements at enabling better machine learning algorithm performance and at providing physical insight into next generation of machine-controlled imaging systems.

Amey Chaware, Colin L. Cooke, Kanghyun Kim et al.

Recent machine learning techniques have dramatically changed how we process digital images. However, the way in which we capture images is still largely driven by human intuition and experience. This restriction is in part due to the many available degrees of freedom that alter the image acquisition process (lens focus, exposure, filtering, etc). Here we focus on one such degree of freedom - illumination within a microscope - which can drastically alter information captured by the image sensor. We present a reinforcement learning system that adaptively explores optimal patterns to illuminate specimens for immediate classification. The agent uses a recurrent latent space to encode a large set of variably-illuminated samples and illumination patterns. We train our agent using a reward that balances classification confidence with image acquisition cost. By synthesizing knowledge over multiple snapshots, the agent can classify on the basis of all previous images with higher accuracy than from naively illuminated images, thus demonstrating a smarter way to physically capture task-specific information.