ABSTRACT
Measuring behavior in the form of numerical data is difficult, especially for studies involving complex actions. DanioVision is a closed-chamber system that utilizes subject tracking to comprehensively record behavior, while also mitigating the influence of environmental conditions. We used DanioVision to record activity of juvenile dwarf cuttlefish (Sepia bandensis) during the inaccessible prey (IP) procedure, a memory experiment in which cuttlefish learn to inhibit capture attempts towards inaccessible prey. By quantifying total movement and orientation of the body, we found that cuttlefish show memory by selectively inhibiting tentacle strikes without reducing total movement, or orientation towards the prey. We show that DanioVision can be used to assess multiple components of dynamic responses that are not measurable by direct observation alone and provide new evidence that strike inhibition is the product of learning, and not motor fatigue.
Subject(s)
Sepia , Animals , Decapodiformes , Learning , Memory , Predatory BehaviorABSTRACT
Microcavity surface plasmon resonance sensors (MSPRSs) develop out of the classic surface plasmon resonance technologies and aim at producing novel lab-on-a-chip devices. MSPRSs generate a series of spectral resonances sensitive to minute changes in the refractive index. Related sensitivity studies and biosensing applications are published elsewhere. The goal of this work is to test the hypothesis that MSPRS resonances are standing surface plasmon waves excited at the surface of the sensor that decay back into propagating photons. Their optical properties (mean wavelength, peak width, and peak intensity) appear highly dependent on the internal morphology of the sensor and the underlying subwavelength aperture architecture in particular. Numerous optical experiments were designed to investigate trends that confirm this hypothesis. An extensive study of prior works was supportive of our findings and interpretations. A complete understanding of those mechanisms and parameters driving the formations of the MSPRS resonances would allow further improvement in sensor sensitivity, reliability, and manufacturability.
ABSTRACT
Personalized medicine has the potential to improve our ability to maintain health and treat disease, while ameliorating continuously rising healthcare costs. Translation of basic research findings to clinical applications within regulatory compliance is required for personalized medicine to become the new foundation for practice of medicine. Deploying even a few of the thousands of potential diagnostic biomarkers identified each year as part of personalized treatment workflows requires clinically efficient biosensor technologies to monitor multiple biomarkers in patients in real time. This paper discusses a critical component of a regulatory system, a microcavity optical biosensor for label-free monitoring of biomolecular interactions at physiologically-relevant concentrations. While most current biosensor research focuses on improving sensitivity, this paper emphasizes other characteristics a biosensor technology requires to be practical in a clinical setting, presenting robust microcavity biosensors which are easy to manufacture and integrate with microfluidics into flexible and redesignable platforms making the microcavity biosensors deployable for continuous monitoring of biomarkers in body fluids in the clinic, in dense 2D random arrays for high-throughput applications like drug-library screening in interactomics, and of the secretory behavior of single cells in the laboratory.
Subject(s)
Biomarkers/chemistry , Biosensing Techniques , Precision Medicine , Humans , Microfluidics , Protein Interaction Maps , Single-Cell Analysis , Surface Plasmon ResonanceABSTRACT
We have developed miniature (approximately 1 microm diameter) microcavity surface-plasmon-resonance sensors (MSPRS), integrated them with microfluidics, and tested their sensitivity to refractive-index changes. We tested their biosensing capability by distinguishing the interaction of glucose oxidase (M(r) 160 kDa) with its natural substrate (beta-D-glucose, M(r) 180 Da) from its interactions with nonspecific substrates (L-glucose, D-mannose, and 2-deoxy-D-glucose). We ran the identical protocol we had used with the MSPRS on a Biacore 3000 instrument using their bare gold chip. Only the MSPRS was able to detect beta-D-glucose binding to glucose oxidase. Each MSPRS can detect the binding to its surface of fewer than 35,000 glucose oxidase molecules (representing 9.6 fg or 60 zmol of protein), about 10(6) times fewer than classical surface-plasmon-resonance biosensors.
Subject(s)
Electrochemical Techniques/instrumentation , Glucose Oxidase/metabolism , Microfluidic Analytical Techniques/instrumentation , Surface Plasmon Resonance/instrumentation , Surface Plasmon Resonance/methods , Electrochemical Techniques/methods , Glucose Oxidase/chemistry , Protein BindingABSTRACT
A miniaturized spherical surface plasmon sensor for measuring the binding kinetics of unlabeled molecules is introduced. The sensor has a submicrometer footprint with a sensitivity that rivals that of state-of-the-art commercial planar surface plasmon sensors, which makes it valuable for applications requiring integration of detection of molecular species in microfluidic channels. The basic principle of the sensor is exploiting the wavelength shifts of the cavity resonances of a metal-coated submicrometer sphere embedded in an opaque metal film due to molecular adsorption. The sensor has been found to be exquisitely responsive in air to water and ethanol vapor adsorption on the bare gold sensor surface. When immersed in a liquid, the sensor can detect the adsorption of less than one monolayer of dodecanethiol (approximately 1.5 nm) on the gold coating of the sphere.
ABSTRACT
We present an improved microfluidic design for generating spatial and temporal gradients. The basic functional elements are bifurcated and trifurcated channels used to split flow between two and three channels, respectively. We use bifurcated channels on the exterior of the channel manifold and trifurcated channels in the interior with mixing tees to recombine flows. For N gradient-forming levels, the number of discrete steps in the gradient is 2(N) + 1, allowing a compact gradient-forming structure that is only 1.6 mm long and 0.5 mm wide. Control of the relative sample concentration at the inlets enables generation of gradients with varying slopes and offsets. The small total channel length allows faster switching (only 2.6 s) between gradients of different compositions than did previous designs, allowing complex temporal sequences and reducing total displacement volume and reagent use. The design permits opposing-gradient experiments and generation of complex nonlinear gradients. We fabricated and tested three channel designs with either three or four gradient-forming levels, 20- or 40-microm channel widths, 60- or 120-microm center-to-center channel spacings, and 9 or 17 output steps. These devices produced essentially identical high-quality linear gradients using both pressure-driven and electrokinetic flow.
Subject(s)
Microfluidic Analytical Techniques/instrumentation , Equipment DesignABSTRACT
Sperm chemotaxis is an area of significant interest to scientists involved in reproductive science. Understanding how and when sperm cells are attracted to the egg could have profound effects on reproduction and contraception. In an effort to systematically study this problem, we have fabricated and evaluated a microfluidic device to measure sperm chemotaxis. The device was designed with a flow-through configuration using a spatially and temporally stable chemical gradient. Mouse sperm cells were introduced into the chemotaxis chamber between confluent flows of mouse ovary extract and buffer. The sperm experiencing chemotaxis swam toward the extract and were counted relative to those that swam toward the buffer. The ovary extracts were diluted from 10(2) to 10(7) times, and each extract dilution was screened for chemotaxis. Four out of six ovaries showed a strong chemotactic response at extract dilutions of 10(-3) to 10(-5). This device provided a convenient, disposable platform on which to conduct chemotaxis assays, and the flow-through design overcomes difficulties associated with distinguishing chemotaxis from trapping.
Subject(s)
Chemotaxis , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods , Spermatozoa/cytology , Animals , Male , Mice , Mice, Inbred ICRABSTRACT
A general method to map the 3D spatial distribution of light emerging from nanoscale apertures is presented that uses photolithographic techniques to create polymer replicas of the intensity distribution. The resulting features varied with aperture diameter and exposure time and showed good correlation with theory. This method provides direct visualization of the intensity distribution in close proximity to nanostructures and overcomes limitations imposed by physical probes where the contribution of the probe to the map requires deconvolution.