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Register a new userPhase Imaging with Computational Specificity (PICS) for measuring dry mass changes in sub-cellular compartments
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Due to its specificity, fluorescence microscopy (FM) has become a quintessential imaging tool in cell biology. However, photobleaching, phototoxicity, and related artifacts continue to limit FMs utility. Recently, it has been shown that artificial intelligence (AI) can transform one form of contrast into another. We present PICS, a combination of quantitative phase imaging and AI, which provides information about unlabeled live cells with high specificity. Our imaging system allows for automatic training, while inference is built into the acquisition software and runs in real-time. Applying the computed fluorescence maps back to the QPI data, we measured the growth of both nuclei and cytoplasm independently, over many days, without loss of viability. Using a QPI method that suppresses multiple scattering, we measured the dry mass content of individual cell nuclei within spheroids. In its current implementation, PICS offers a versatile quantitative technique for continuous simultaneous monitoring of individual cellular components in biological applications where long-term label-free imaging is desirable.
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Quantum correlation and its measurement are essential in exploring fundamental quantum physics problems and developing quantum enhanced technologies. Quantum correlation may be generated and manipulated in different spaces, which demands different measurement approaches corresponding to position, time, frequency and polarization of quantum particles. In addition, after early proof-of-principle demonstrations, it is of great demand to measure quantum correlation in a Hilbert space large enough for real quantum applications. When the number of modes goes up to several hundreds, it becomes economically unfeasible for single-mode addressing and also extremely challenging for processing correlation events with hardware. Here we present a general and large-scale measurement approach of Correlation on Spatially-mapped Photon-Level Image (COSPLI). The quantum correlations in other spaces are mapped into the position space and are captured by single-photon-sensitive imaging system. Synthetic methods are developed to suppress noises so that single-photon registrations can be faithfully identified in images. We eventually succeed in retrieving all the correlations with big-data technique from tens of millions of images. We demonstrate our COSPLI by measuring the joint spectrum of parametric down-conversion photons. Our approach provides an elegant way to observe the evolution results of large-scale quantum systems, representing an innovative and powerful tool added into the platform for boosting quantum information processing.
Fluorescence microscopy is a powerful tool to measure molecular specific information in biological samples. However, most biological tissues are highly heterogeneous because of refractive index (RI) differences and thus degrade the signal-to-noise ratio of fluorescence images. At the same time, RI is an intrinsic optical property of label free biological tissues that quantitatively relates to cell morphology, mass, and stiffness. Conventional imaging techniques measure fluorescence and RI of biological samples separately. Here, we develop a new computational hybrid imaging method based on a multi-slice model of multiple scattering that reconstructs 3D fluorescence and 3D RI from the same dataset of fluorescence images. Our method not only bridges the gap between fluorescence and RI imaging and provides a panoramic view of the biological samples, but also can digitally correct multiple scattering effect of fluorescence images from the reconstructed 3D RI. Computational hybrid imaging opens a unique avenue beyond conventional imaging techniques.
Label-free imaging approaches seek to simplify and augment histopathologic assessment by replacing the current practice of staining by dyes to visualize tissue morphology with quantitative optical measurements. Quantitative phase imaging (QPI) operates with visible/UV light and thus provides a resolution matched to current practice. Here we introduce and demonstrate confocal QPI for label-free imaging of tissue sections and assess its utility for manual histopathologic inspection. Imaging cancerous and normal adjacent human breast and prostate, we show that tissue structural organization can be resolved with high spatial detail comparable to conventional H&E stains. Our confocal QPI images are found to be free of halo, solving this common problem in QPI. We further describe and apply a virtual imaging system based on Finite-Difference Time-Domain (FDTD) calculations to quantitatively compare confocal with wide-field QPI methods and explore performance limits using numerical tissue phantoms.
The ability to acquire large-scale recordings of neuronal activity in awake and unrestrained animals poses a major challenge for studying neural coding of animal behavior. We present a new instrument capable of recording intracellular calcium transients from every neuron in the head of a freely behaving C. elegans with cellular resolution while simultaneously recording the animals position, posture and locomotion. We employ spinning-disk confocal microscopy to capture 3D volumetric fluorescent images of neurons expressing the calcium indicator GCaMP6s at 5 head-volumes per second. Two cameras simultaneously monitor the animals position and orientation. Custom software tracks the 3D position of the animals head in real-time and adjusts a motorized stage to keep it within the field of view as the animal roams freely. We observe calcium transients from 78 neurons and correlate this activity with the animals behavior. Across worms, multiple neurons show significant correlations with modes of behavior corresponding to forward, backward, and turning locomotion. By comparing the 3D positions of these neurons with a known atlas, our results are consistent with previous single-neuron studies and demonstrate the existence of new candidate neurons for behavioral circuits.
Aging affects almost all aspects of an organism -- its morphology, its physiology, its behavior. Isolating which biological mechanisms are regulating these changes, however, has proven difficult, potentially due to our inability to characterize the full repertoire of an animals behavior across the lifespan. Using data from fruit flies (D. melanogaster) we measure the full repertoire of behaviors as a function of age. We observe a sexually dimorphic pattern of changes in the behavioral repertoire during aging. Although the stereotypy of the behaviors and the complexity of the repertoire overall remains relatively unchanged, we find evidence that the observed alterations in behavior can be explained by changing the flys overall energy budget, suggesting potential connections between metabolism, aging, and behavior.
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