Drug discovery and development are hampered by high failure rates attributed to the reliance on non-human animal models employed during safety and efficacy testing. A fundamental problem in this inefficient process is that non-human animal models cannot adequately represent human biology. Thus, there is an urgent need for high-content in vitro systems that can better predict drug-induced toxicity. Systems that predict cardiotoxicity are of uppermost significance, as approximately one third of safety-based pharmaceutical withdrawals are due to cardiotoxicty. Here, we present a cardiac microphysiological system (MPS) with the attributes required for an ideal in vitro system to predict cardiotoxicity: i) cells with a human genetic background; ii) physiologically relevant tissue structure (e.g. aligned cells); iii) computationally predictable perfusion mimicking human vasculature; and, iv) multiple modes of analysis (e.g. biological, electrophysiological, and physiological). Our MPS is able to keep human induced pluripotent stem cell derived cardiac tissue viable and functional over multiple weeks. Pharmacological studies using the cardiac MPS show half maximal inhibitory/effective concentration values (IC50/EC50) that are more consistent with the data on tissue scale references compared to cellular scale studies. We anticipate the widespread adoption of MPSs for drug screening and disease modeling.

Dravet syndrome is a catastrophic childhood epilepsy with early-onset seizures, delayed language and motor development, sleep disturbances, anxiety-like behaviour, severe cognitive deficit and an increased risk of fatality. It is primarily caused by de novo mutations of the SCN1A gene encoding a neuronal voltage-activated sodium channel. Zebrafish with a mutation in the SCN1A homologue recapitulate spontaneous seizure activity and mimic the convulsive behavioural movements observed in Dravet syndrome. Here, we show that phenotypic screening of drug libraries in zebrafish scn1 mutants rapidly and successfully identifies new therapeutics. We demonstrate that clemizole binds to serotonin receptors and its antiepileptic activity can be mimicked by drugs acting on serotonin signalling pathways e.g. trazodone and lorcaserin. Coincident with these zebrafish findings, we treated five medically intractable Dravet syndrome patients with a clinically-approved serotonin receptor agonist (lorcaserin, Belviq®) and observed some promising results in terms of reductions in seizure frequency and/or severity. Our findings demonstrate a rapid path from preclinical discovery in zebrafish, through target identification, to potential clinical treatments for Dravet syndrome.

Nucleic acid amplification and quantification via polymerase chain reaction (PCR) is one of the most sensitive and powerful tools for clinical laboratories, precision medicine, personalized medicine, agricultural science, forensic science and environmental science. Ultrafast multiplex PCR, characterized by low power consumption, compact size and simple operation, is ideal for timely diagnosis at the point-of-care (POC). Although several fast/ultrafast PCR methods have been proposed, the use of a simple and robust PCR thermal cycler remains challenging for POC testing. Here, we present an ultrafast photonic PCR method using plasmonic photothermal light-to-heat conversion via photon-electron-phonon coupling. We demonstrate an efficient photonic heat converter using a thin gold (Au) film due to its plasmon-assisted high optical absorption (approximately 65% at 450 nm, the peak wavelength of heat source light-emitting diodes (LEDs)). The plasmon-excited Au film is capable of rapidly heating the surrounding solution to over 150 6C within 3 min. Using this method, ultrafast thermal cycling (30 cycles; heating and cooling rate of 12.7960.93 6C s 21 and 6.660.29 6C s 21 , respectively) from 55 6C (temperature of annealing) to 95 6C (temperature of denaturation) is accomplished within 5 min. Using photonic PCR thermal cycles, we demonstrate here successful nucleic acid (l-DNA) amplification. Our simple, robust and low cost approach to ultrafast PCR using an efficient photonic-based heating procedure could be generally integrated into a variety of devices or procedures, including on-chip thermal lysis and heating for isothermal amplifications.

Nanocorals demonstrate independent cell‐targeting and label‐free biomolecular sensing capabilities. The highly roughened gold region of the nanocoral increases the adsorption capacity and causes a strong surface‐enhanced Raman spectroscopy (SERS) signal. The polystyrene region can adsorb antibodies, allowing the nanocoral to specifically bind to receptors on the cancer cell membrane making nanocorals multifunctional nanosensors for cellular diagnostics and treatment evaluation.

A real-time label-free detection method is in critical demand for biological, chemical, and medical applications, and environmental monitoring. Surface-enhanced Raman spectroscopy (SERS) is one of the best label-free detection methods. [1][2][3] Accordingly, a variety of fabrication methods of SERS substrates have been demonstrated. [4][5][6] Among the many such fabrication methods, self-assembly of nanoparticles is dominant. However, due to the uncontrollable bottom-up approach, the self-assembly method forms random hot spots for electromagnetic (EM) field enhancement. [6] Since the disordered random hot spots cause a nonuniform response of SERS signals, it is required to find a solution for forming precisely predetermined, reproducible, and organized nanoarchitectures for SERS substrates. For these purposes, focused-ion-beam [7,8] and electron-beam lithography [9] were applied for precision nanopatterning with the advantage of high resolution without the need for a physical mask, since the pattern can be changed at any time by using computer-aided design (CAD) software. However, the disadvantages of these two methods are the long exposure time due to pixel-by-pixel scanning steps, high cost, and substantial maintenance. To overcome these limitations, phase-shift lithography, [10] nanosphere lithography, [11] andcommunications Figure 5. Uniform SERS spectra on the nanopore SERS array. The spectra were measured at 15 different positions on a 4 cm 2 area of AAO coated with 20 nm Au; t AAO ¼ 100 nm and D nc ¼ 32 nm. Variation in the SERS spectra was well below 10%.

A cell in vivo is part of a large, networked community. An individual cell's fate is strongly influenced by its interactions with neighbouring cells. While this interaction has been recognized as critical in determining cell behaviour, its complexity and heterogeneity has thus far defied characterization by currently available techniques. Herein, we present a single-cell level co-culture platform for studies of dynamic cellular interactions, which is capable of maintaining and tracking single-cell pair interactions to simplify the complexity of intercellular communication. In this platform, heterotypic pairing on a single-cell level is achieved through sequential cell trapping and dynamic variation of fluidic resistance. Individual culture chambers provide trapped cells enough space to migrate and proliferate through multiple generations. Furthermore, the semi-isolated chambers, combined with continuously refreshed medium supplement, allow a stable communication environment around the cells. To demonstrate the platform capability, we cultured and tracked stem cell-fibroblast pairs for several generations. The subsequent effects of cell-cell interactions were then easily observed, due to the addressability of each isolated chamber, and quantitatively characterized. Specifically, we found that paired cells' migration patterns were dependent on their initial distance from each other, and that heterotypic pairing led to distinct proliferation patterns from homotypic, single-cell culture. This study demonstrates the platform utility in providing a detailed and quantitative understanding of the complexity of cellular communication and its effects on cell behaviour in a variety of biological systems.

Oriented assemblies of functional nanoparticles, with the aid of external physical and chemical driving forces, have been prepared on two-dimensional solid substrates. It is challengeable, however, to achieve three-dimensional assembly directly in solution, owing to thermal fluctuations and free diffusion. Here we describe the self-orientation of gold nanorods at an immiscible liquid interface (that is, oleic acid-water) and exploit this novel phenomenon to create a substrate-free interfacial liquid-state surface-enhanced Raman spectroscopy. Dark-field imaging and Raman scattering results reveal that gold nanorods spontaneously adopt a vertical orientation at an oleic acid-water interface in a stable trapping mode, which is in good agreement with simulation results. The spontaneous vertical alignment of gold nanorods at the interface allows one to accomplish significant additional amplification of the Raman signal, which is up to three to four orders of magnitude higher than that from a solution of randomly oriented gold nanorods.

Epilepsy is a common chronic neurological disease affecting almost 3 million people in the United States and 50 million people worldwide. Despite availability of more than two dozen FDA-approved anti-epileptic drugs (AEDs), one-third of patients fail to receive adequate seizure control. Specifically, pediatric genetic epilepsies are often the most severe, debilitating and pharmaco-resistant forms of epilepsy. Epileptic syndromes share a common symptom of unprovoked seizures. While some epilepsies/forms of epilepsy are the result of acquired insults such as head trauma, febrile seizure, or viral infection, others have a genetic basis. The discovery of epilepsy associated genes suggests varied underlying pathologies and opens the door for development of new “personalized” treatment options for each genetic epilepsy. Among these, Dravet syndrome (DS) has received substantial attention for both the pre-clinical and early clinical development of novel therapeutics. Despite these advances, there is no FDA-approved treatment for DS. Over 80% of patients diagnosed with DS carry a de novo mutation within the voltage-gated sodium channel gene SCN1A and these patients suffer with drug resistant and life-threatening seizures. Here we will review the preclinical animal models for DS featuring inactivation of SCN1A (including zebrafish and mice) with an emphasis on seizure phenotypes and behavioral comorbidities. Because many drugs fail somewhere between initial preclinical discovery and clinical trials, it is equally important that we understand how these models respond to known AEDs. As such, we will also review the available literature and recent drug screening efforts using these models with a focus on assay protocols and predictive pharmacological profiles. Validation of these preclinical models is a critical step in our efforts to efficiently discover new therapies for these patients. The behavioral and electrophysiological drug screening assays in zebrafish will be discussed in detail including specific examples from our laboratory using a zebrafish scn1 mutant and a summary of the nearly 3000 drugs screened to date. As the discovery and development phase rapidly moves from the lab-to-the-clinic for DS, it is hoped that this preclinical strategy offers a platform for how to approach any genetic epilepsy.