Abstract:To study the behavior of freely moving model organisms such as zebrafish (Danio rerio) and fruit flies (Drosophila) across multiple spatial scales, it would be ideal to use a light microscope that can resolve 3D information over a wide field of view (FOV) at high speed and high spatial resolution. However, it is challenging to design an optical instrument to achieve all of these properties simultaneously. Existing techniques for large-FOV microscopic imaging and for 3D image measurement typically require many sequential image snapshots, thus compromising speed and throughput. Here, we present 3D-RAPID, a computational microscope based on a synchronized array of 54 cameras that can capture high-speed 3D topographic videos over a 135-cm^2 area, achieving up to 230 frames per second at throughputs exceeding 5 gigapixels (GPs) per second. 3D-RAPID features a 3D reconstruction algorithm that, for each synchronized temporal snapshot, simultaneously fuses all 54 images seamlessly into a globally-consistent composite that includes a coregistered 3D height map. The self-supervised 3D reconstruction algorithm itself trains a spatiotemporally-compressed convolutional neural network (CNN) that maps raw photometric images to 3D topography, using stereo overlap redundancy and ray-propagation physics as the only supervision mechanism. As a result, our end-to-end 3D reconstruction algorithm is robust to generalization errors and scales to arbitrarily long videos from arbitrarily sized camera arrays. The scalable hardware and software design of 3D-RAPID addresses a longstanding problem in the field of behavioral imaging, enabling parallelized 3D observation of large collections of freely moving organisms at high spatiotemporal throughputs, which we demonstrate in ants (Pogonomyrmex barbatus), fruit flies, and zebrafish larvae.
Abstract: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.
Abstract: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.
Abstract: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.