Recent advances in microfluidics to generate and control picoliter emulsions of water in oil have enabled ultra-sensitive assays for small molecules, proteins, nucleic acids, and cells. Unfortunately, the conventional fluorescence detection used to measure the outcome of these droplet-based assays has not proven suited to match the time and space multiplexing capabilities of microfluidic systems. To address this challenge, we developed an in-flow fluorescence detection platform that enables multiple streams of droplets to be monitored using only a single photodetector and no lenses. The key innovation of our technology is the amplitude modulation of the signal from fluorescent droplets using distinct micro-patterned masks for each channel. By taking advantage of the high bandwidth of electronics, our technique enables the velocity-independent recovery of weak fluorescent signals (SNR ≪ 1) using only simple hardware, obviating the need for lasers, bulky detectors, and complex fluid control. We demonstrated a handheld-sized device that simultaneously monitors four independent channels with the capability to be scaled-up to more than sixteen, limited primarily by the droplet density.

The goal of this study is to identify the most fundamental independent elements common to natural color images that can be applied for general image representation, and examine their possible neurophysiological correlates. To this end we applied Independent Component Analysis (ICA) to naturally occurring color images. ICA derives statistically independent image elements by minimizing mutual information. We used images consisting of flowers, leaves, trees, rocks, and other natural objects and background. ICA results in luminance and color filters with simple and complex cell receptive field profiles. The luminance filters are localized and oriented edge detectors as reported previously (1). The color filters comprise of blue-yellow and red-green double-opponent receptive fields with various orientations. ICA shows that the independent components of natural color images are multispectral edges, and predicts that spatio-chromatic information is coded in statistically independent luminance, blue-yellow, and red-green opponent pathways. The color and luminance receptive fields may be considered optimal for edge and orientation detection in color or multispectral images (2). Despite the lack of strong physiological evidence for double-opponent cells in mammalian cortex, ICA supports that decomposition of spatio-chromatic information into luminance, red-green, and blue-yellow channels offers an optimum means of coding natural images. A possibility is that the local computation of double opponency is not carried out by individual cells but distributed across a population of cells. (1) BellA. J.SejnowskiT. J.Vision Research, 37, 3327–3338, 1997 (2) TailorD.FinkelL. H.BuchsbaumG.Vision Research, 40, 2671–2676, 2000

Non-negative matrix factorization (NMF, Nature 401 (1999) 788-791) is a method to derive non-negative basis functions for sets of data that are inherently non-negative, such as color spectra. We applied NMF to Munsell color spectra and investigated the color names associated with the non-negative basis functions. NMF yields basis functions compatible with established color naming categories.

Cone synaptic terminals couple electrically to their neighbors. This reduces the amplitude of temporally uncorrelated voltage differences between neighbors. For an achromatic stimulus coarser than the cone mosaic, the uncorrelated voltage difference between neighbors represents mostly noise; so noise is reduced more than the signal. Here coupling improves signal-to-noise ratio and enhances contrast sensitivity. But for a chromatic stimulus the uncorrelated voltage difference between neighbors of different spectral type represents mostly signal; so signal would be reduced more than the noise. This cost of cone coupling to encoding chromatic signals was evaluated using a compartmental model of the foveal cone array. When cones sensitive to middle (M) and long (L) wavelengths alternated regularly, and the conductance between a cone and all of its immediate neighbors was 1000 pS (similar to 2 connexons/cone pair), coupling reduced the difference between the L and M action spectra by nearly fivefold, from about 38% to 8%. However, L and M cones distribute randomly in the mosaic, forming small patches of like type, and within a patch the responses to a chromatic stimulus are correlated. In such a mosaic, coupling still reduced the difference between the I, and M action spectra, but only by 2.4-fold, to about 18%. This result is independent of the L/M ratio. Thus "patchiness" of the L/M mosaic allows cone coupling to improve achromatic contrast sensitivity while minimizing the cost to chromatic sensitivity. (C) 2000 Optical Society of America [S0740-3232(00)02103-7].

Independent Component Analysis (ICA) of images of natural scenes has been shown to generate basis functions, or filters, which resemble spatial [Bell & Sejnowski (1997). Vision Research, 37, 3327-3338; van Hateren & van der Schaaf (1998). Proceedings of the Royal Society of London B, 265, 359-366] and spatiotemporal [van Hateren & Ruderman (1998) Proceedings of the Royal Society of London B, 265, 2315-2320] receptive fields of simple cells of the striate cortex. ICA yields statistically independent components which provide for a redundancy-reduced representation of the data. Using one of several published algorithms [Lee (1998). Independent component analysis: theory and applications. Boston; Kluwer Academic], we applied linear ICA to color images of natural scenes. The resulting independent component filters (ICFs) separate into either luminance or color filters. The luminance filters are localized and oriented edge detectors as reported previously. The color filters resemble either blue-yellow or red-green double-opponent receptive fields with various orientations. An equal number of each type of filter (luminance, red-green, and blue-yellow) is obtained. Thus, ICA predicts that spatiochromatic information is coded in statistically independent luminance, blue-yellow, and red-green opponent pathways with a relatively equal representation and specific spatial profiles at the cortical level.

Transmitter concentration at a three-dimensional synapse. J. Neurophysiol. 80: 3163-3172, 1998. At intensities from starlight to 1000-fold brighter, the mammalian rod synapse transmits a binary signal, the capture of 0 or 1 photon. Zero is signified by tonic exocytosis, and 1 is signified by a brief pause. The synapse is three dimensional: vesicles discharge at the apex of a deep cleft created by the invagination of four postsynaptic processes. Two horizontal cell spines bearing alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors reach near to the release sites (16 nm), and two bipolar dendrites bearing mGluR6 receptors end far from the release sites (up to 640 nm). We considered two hypotheses for signal transfer: transmitter quanta might be integrated in the cleft and sensed as a steady concentration (high for 0 and low for 1); or quanta might be sensed at the postsynaptic membrane as discrete postsynaptic potentials (PSPs) and integrated within the dendrite. We calculate from a passive diffusion model that the invagination empties rapidly (tau approximately 1.7 ms). Further calculations suggest that a glutamate concentration high enough to hold a bipolar cell in darkness at one end of its response range would require approximately 4,000 vesicles/s. On the other hand, the glutamate pulse from a single vesicle would reach both nearby AMPA receptors (low affinity) and distant mGluR6 receptors (high affinity) at spatiotemporal concentrations matched to their apparent binding affinities. Thus one vesicle could evoke a discrete PSP in all four postsynaptic processes. We calculate from a stochastic model that PSPs could transfer the binary signal at approximately 100 vesicles/s. Thus dendritic integration of unitary PSPs is both plausible and 40-fold more efficient than the alternative mechanism. The rod's deep invagination, rather than serving to pool transmitter, may serve to prevent "spillover" of transmitter to neighboring rods. Spillover, by pooling the noise from neighboring rods, would impair transmission of their binary signals.

An important issue in understanding the retina is finding candidate functional roles for different cell pathways and the details of their anatomy and physiology. We consider various spatial properties of the three main cone ==> cone bipolar cell ==> on-beta ganglion cell pathways in the cat retina and possible roles for the particulars of their anatomy. The cone bipolar cells in these pathways have distinct morphologies and modest differences in their convergence, divergence, densities, and synaptic weighting; and it is unclear whether the pathways differ in their spatial properties or in some other manner. Since differences in spatial processing of cells are best studied on a systemwide level, we developed the multirate filter-based method of retinal modeling, a technique for relating the anatomy of multiple cell layers to its systemic effects. We demonstrate that (1) despite the anatomic distinctions among the three main cone bipolar cell pathways, their spatial responses are essentially identical; (2) despite the spatial averaging in the pathways, there is essentially no filtering of the nonaliasing signal components after the cone layer; (3) instead, this averaging combined with prefiltering by the eye's optics and cone gap junctions prevents spatial aliasing; and (4) the averaging and prefiltering combined allow cell responses to be similar despite significant cell-to-cell anatomic differences.

Invariance of cone ratios has been proposed to be a possible basis for perceptual color constancy. Cone excitation ratios were computed across reflectance edges viewed under different illuminants as well as across illuminant edges. The results of the authors' computations suggest that such a mechanism is incomplete. Cone ratios maintain color constancy across reflectance edges under a variety of illuminants, yet they are unable to discern whether a border is a reflectance edge or an illuminant edge

A biologically-based multistage neural network is presented which produces color constant responses to a variety of color stimuli. The network takes advantage of several mechanisms in the human visual system, including retinal adaptation, spectral opponency, and spectrally-specific long-range inhibition. This last stage is a novel mechanism based on cells which have been described in cortical area V4. All stages include nonlinear response functions. The model emulates human performance in several psychophysical paradigms designed to test color constancy and color induction. We measured the amount of constancy achieved with both natural and artificial simulated illuminants, using homogeneous grey backgrounds and more complex backgrounds, such as Mondrians. On average, the model performs as well or better than the average human color constancy performance under similar conditions. The network simulation also displays color induction and assimilation behavior consistent with human perceptual data.

Abstract : Color constancy, or the ability of the visual system to perceive color independently of the ambient illumination, was investigated in the context of a biologically-based neural network. In particular, the role of retinal adaptation and higher level visual operations in mediating color constancy was investigated. The study incorporated properties of individual cells and how they combine to make complex color and spatial operations. The neural network simulations indicate how early visual stages complement each other to compensate and maintain relatively constant color perception under conditions of varying illumination and spatial context in the image. The network takes advantage of several mechanisms in the human visual system, including retinal adaptation, spectral opponency, and spectrally-specific long-range inhibition. This last stage is a novel mechanism based on cells which have been described in cortical area v4. All stages include non-linear response functions. The model emulates human performance in several psychophysical paradigms designed to test color constancy and color induction. We measured the amount of constancy achieved with both natural and artificial simulated illuminants, using homogeneous gray backgrounds and more complex backgrounds, such as Mondrians.

The mammalian rod synapse transmits a binary signal (one photon or none) using tonic, rapid exocytosis. We constructed a quantitative, physical model of the synapse. Presynaptically, a single, linear active zone provides docking sites for approximately 130 vesicles, and a "ribbon" anchored to the active zone provides a depot for approximately 640 vesicles. Postsynaptically, 4 processes invaginate the terminal: 2 (known to have low affinity glutamate receptors) lie near the active zone (16 nm), and 2 (known to have high affinity glutamate receptors) lie at a distance (130-640 nm). The presynaptic structure seems designed to minimize fluctuations in tonic rate owing to empty docking sites, whereas the postsynaptic geometry may permit 1 vesicle to evoke an all-or-none response at all 4 postsynaptic processes.

A biologically-based neural network simulation is used to analyze the contributions to color perception of each of several processing steps in the visual system from the retina to cortical area V4. We consider the effects on color constancy and color induction of adaptation, spectral opponency, non-linearities including saturation and rectification, and spectrally-specific long-range inhibition. This last stage is a novel mechanism based on cells which have been described in V4. The model has been tested with simulations of several well known psychophysical color constancy and color induction experiments. We conclude from these simulations the following: (1) a simple push-pull spectrally specific contrast mechanism, using large surrounds analogous to those found in V4, is very effective in producing general color constancy and color induction behavior; (2) given some spatio-temporal averaging, receptor adaptation can also produce a degree of color constancy; (3) spectrally opponent processes have spatial frequency dependent responses to color and brightness contrast which affect the contribution of the V4 mechanism to color constancy in images with nonuniform backgrounds; and (4) the effect of the V4 mechanism depends on the difference between center and surround while the effect of adaptation depends on the total sum of inputs from both center and surround and therefore the two stages cooperate to increase the range of stimulus conditions under which color constancy can be achieved.

Under scotopic conditions, the mammalian rod encodes either one photon or none within its integration time. Consequently the signal presented to its synaptic terminal is binary. The synapse has a single active zone that releases neurotransmitter quanta tonically in darkness and pauses briefly in response to a rhodopsin isomerization by a photon. We asked: what minimum tonic rate would allow the postsynaptic bipolar cell to distinguish this pause from an extra-long interval between quanta due to the stochastic timing of release? The answer required a model of the circuit that included the rod convergence onto the bipolar cell and the bipolar cell's signal-to-noise ratio. Calculations from the model suggest that tonic release must be at least 40 quanta/s. This tonic rate is much higher than at conventional synapses where reliability is achieved by employing multiple active zones. The rod's synaptic mechanism makes efficient use of space, which in the retina is at a premium.

The viscoelastic properties of the vitreous body from bovine and porcine eyes were determined by microrheometry. Each vitreous sample was sectioned into anterior, central and posterior segments. The rheological properties of each species and region were compared with each other and with the viscoelastic properties of human vitreous reported earlier. The results showed significant variations among species, as well as between regions in all species. All regions of human vitreous have significantly different rheological properties from those of the cow, and from the anterior and posterior regions of the pig; however, the viscoelastic behavior of the central porcine region closely resembles that of the human. In comparing the three species, the major differences found are in those parameters characterizing ''viscous'' or dissipative properties; values of ''elastic'' or energy storage properties were similar.