SynQuant: an automatic tool to quantify synapses from microscopy images.

MOTIVATION: Synapses are essential to neural signal transmission. Therefore, quantification of synapses and related neurites from images is vital to gain insights into the underlying pathways of brain functionality and diseases. Despite the wide availability of synaptic punctum imaging data, several issues are impeding satisfactory quantification of these structures by current tools. First, the antibodies used for labeling synapses are not perfectly specific to synapses. These antibodies may exist in neurites or other cell compartments. Second, the brightness of different neurites and synaptic puncta is heterogeneous due to the variation of antibody concentration and synapse-intrinsic differences. Third, images often have low signal to noise ratio due to constraints of experiment facilities and availability of sensitive antibodies. These issues make the detection of synapses challenging and necessitates developing a new tool to easily and accurately quantify synapses. RESULTS: We present an automatic probability-principled synapse detection algorithm and integrate it into our synapse quantification tool SynQuant. Derived from the theory of order statistics, our method controls the false discovery rate and improves the power of detecting synapses. SynQuant is unsupervised, works for both 2D and 3D data, and can handle multiple staining channels. Through extensive experiments on one synthetic and three real datasets with ground truth annotation or manually labeling, SynQuant was demonstrated to outperform peer specialized unsupervised synapse detection tools as well as generic spot detection methods. AVAILABILITY AND IMPLEMENTATION: Java source code, Fiji plug-in, and test data are available at https://github.com/yu-lab-vt/SynQuant. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Efficient Global MOT Under Minimum-Cost Circulation Framework.

We developed a minimum-cost circulation framework for solving the global data association problem, which plays a key role in the tracking-by-detection paradigm of multi-object tracking (MOT). The global data association problem was extensively studied under the minimum-cost flow framework, which is theoretically attractive as being flexible and globally solvable. However, the high computational burden has been a long-standing obstacle to its wide adoption in practice. While enjoying the same theoretical advantages and maintaining the same optimal solution as the minimum-cost flow framework, our new framework has a better theoretical complexity bound and leads to orders of practical efficiency improvement. This new framework is motivated by the observation that minimum-cost flow only partially models the data association problem and it must be accompanied by an additional and time-consuming searching scheme to determine the optimal object number. By employing a minimum-cost circulation framework, we eliminate the searching step and naturally integrate the number of objects into the optimization problem. By exploring the special property of the associated graph, that is, an overwhelming majority of the vertices are with unit capacity, we designed an implementation of the framework and proved it has the best theoretical computational complexity so far for the global data association problem. We evaluated our method with 40 experiments on five MOT benchmark datasets. Our method was always the most efficient in every single experiment and averagely 53 to 1,192 times faster than the three state-of-the-art methods. When our method served as a sub-module for global data association methods utilizing higher-order constraints, similar running time improvement was attained. We further illustrated through several case studies how the improved computational efficiency enables more sophisticated tracking models and yields better tracking accuracy. We made the source code publicly available on GitHub with both Python and MATLAB interfaces.

A Neural Basis for Categorizing Sensory Stimuli to Enhance Decision Accuracy.

Sensory stimuli with graded intensities often lead to yes-or-no decisions on whether to respond to the stimuli. How this graded-to-binary conversion is implemented in the central nervous system (CNS) remains poorly understood. Here, we show that graded encodings of noxious stimuli are categorized in a decision-associated CNS region in Drosophila larvae, and then decoded by a group of peptidergic neurons for executing binary escape decisions. GABAergic inhibition gates weak nociceptive encodings from being decoded, whereas escalated amplification through the recruitment of second-order neurons boosts nociceptive encodings at intermediate intensities. These two modulations increase the detection accuracy by reducing responses to negligible stimuli whereas enhancing responses to intense stimuli. Our findings thus unravel a circuit mechanism that underlies accurate detection of harmful stimuli.

Automated Functional Analysis of Astrocytes from Chronic Time-Lapse Calcium Imaging Data.

Recent discoveries that astrocytes exert proactive regulatory effects on neural information processing and that they are deeply involved in normal brain development and disease pathology have stimulated broad interest in understanding astrocyte functional roles in brain circuit. Measuring astrocyte functional status is now technically feasible, due to recent advances in modern microscopy and ultrasensitive cell-type specific genetically encoded Ca2+ indicators for chronic imaging. However, there is a big gap between the capability of generating large dataset via calcium imaging and the availability of sophisticated analytical tools for decoding the astrocyte function. Current practice is essentially manual, which not only limits analysis throughput but also risks introducing bias and missing important information latent in complex, dynamic big data. Here, we report a suite of computational tools, called Functional AStrocyte Phenotyping (FASP), for automatically quantifying the functional status of astrocytes. Considering the complex nature of Ca2+ signaling in astrocytes and low signal to noise ratio, FASP is designed with data-driven and probabilistic principles, to flexibly account for various patterns and to perform robustly with noisy data. In particular, FASP explicitly models signal propagation, which rules out the applicability of tools designed for other types of data. We demonstrate the effectiveness of FASP using extensive synthetic and real data sets. The findings by FASP were verified by manual inspection. FASP also detected signals that were missed by purely manual analysis but could be confirmed by more careful manual examination under the guidance of automatic analysis. All algorithms and the analysis pipeline are packaged into a plugin for Fiji (ImageJ), with the source code freely available online at https://github.com/VTcbil/FASP.