High-throughput optofluidic screening for improved microbial cell factories via real-time micron-scale productivity monitoring.

The industrial synthetic biology sector has made huge investments to achieve relevant miniaturized screening systems for scalable fermentation. Here we present the first example of a high-throughput (>10 genotypes per week) perfusion-based screening system to improve small-molecule secretion from microbial strains. Using the Berkeley Lights Beacon® system, the productivity of each strain could be directly monitored in real time during continuous culture, yielding phenotypes that correlated strongly (r > 0.

Fairness versus efficiency: how procedural fairness concerns affect coordination.

We investigate in a laboratory experiment whether procedural fairness concerns affect how well individuals are able to solve a coordination problem in a two-player Volunteer's Dilemma. Subjects receive external action recommendations, either to volunteer or to abstain from it, in order to facilitate coordination and improve efficiency. We manipulate the fairness of the recommendation procedure by varying the probabilities of receiving the disadvantageous recommendation to volunteer between players.

Synthetic Capillaries to Control Microscopic Blood Flow.

Capillaries pervade human physiology. The mean intercapillary distance is only about 100 μm in human tissue, which indicates the extent of nutrient diffusion. In engineered tissue the lack of capillaries, along with the associated perfusion, is problematic because it leads to hypoxic stress and necrosis.

Single cell transfection with single molecule resolution using a synthetic nanopore.

We report the development of a single cell gene delivery system based on electroporation using a synthetic nanopore, that is not only highly specific and very efficient but also transfects with single molecule resolution at low voltage (1 V) with minimal perturbation to the cell. Such a system can be used to control gene expression with unprecedented precision--no other method offers such capabilities.

Biological noise abatement: coordinating the responses of autonomous bacteria in a synthetic biofilm to a fluctuating environment using a stochastic bistable switch.

Noise is inherent to single cell behavior. Its origins can be traced to the stochasticity associated with a few copies of genes and low concentrations of protein and ligands. We have studied the mechanisms by which the response of noisy elements can be entrained for biological signal processing.

Epigenetic memory emerging from integrated transcription bursts.

Some autonomous bacteria coordinate their actions using quorum-sensing (QS) signals to affect gene expression. However, noise in the gene environment can compromise the cellular response. By exercising precise control over a cell's genes and its microenvironment, we have studied the key positive autoregulation element by which the lux QS system integrates noisy signals into an epigenetic memory.

Direct visualization of single-molecule translocations through synthetic nanopores comparable in size to a molecule.

A nanopore is the ultimate analytical tool. It can be used to detect DNA, RNA, oligonucleotides, and proteins with submolecular sensitivity. This extreme sensitivity is derived from the electric signal associated with the occlusion that develops during the translocation of the analyte across a membrane through a pore immersed in electrolyte.

Using a nanopore for single molecule detection and single cell transfection.

We assert that it is possible to trap and identify proteins, and even (conceivably) manipulate proteins secreted from a single cell (i.e. the secretome) through transfection via electroporation by exploiting the exquisite control over the electrostatic potential available in a nanopore.

Sample cells for probing solid/liquid interfaces with broadband sum-frequency-generation spectroscopy.

Two sample cells designed specifically for sum-frequency-generation (SFG) measurements at the solid/liquid interface were developed: one thin-layer analysis cell allowing measurement of films on reflective metallic surfaces through a micrometer layer of solution and one spectroelectrochemical cell allowing investigation of processes at the indium tin oxide/solution interface. Both sample cells are described in detail and data illustrating the capabilities of each are shown. To further improve measurements at solid/liquid interfaces, the broadband SFG system was modified to include a reference beam which can be measured simultaneously with the sample signal, permitting background correction of SFG spectra in real time.

In situ characterization of thermo-responsive poly(N-isopropylacrylamide) films with sum-frequency generation spectroscopy.

The thermo-responsive behaviour of thiol modified poly(N-isopropylacrylamide) (pNIPAM) films immobilized on gold are probed by in situ broadband sum-frequency generation (SFG) spectroscopy. The pNIPAM films were prepared by atom transfer radical polymerization (ATRP) using a nitro-biphenyl-thiol (NBT)-SAM on a polycrystalline gold surface as a substrate. Additionally, Raman and infrared reflection absorption spectroscopy (IRRAS) are applied to spin-coated pNIPAM films.

Pulse oximeter perfusion index as an early indicator of sympathectomy after epidural anesthesia.

Background: The pulse oximeter perfusion index (PI) has been used to indicate sympathectomy-induced vasodilatation. We hypothesized that pulse oximeter PI provides an earlier and clearer indication of sympathectomy following epidural anesthesia than skin temperature and arterial pressure.

Methods: Forty patients received lumbar epidural catheters.

Sympathectomy-mediated vasodilatation: a randomized concentration ranging study of epidural bupivacaine.

Purpose: We tested the hypothesis that the development of sympathectomy-mediated vasodilatation is dependent on the concentration rather than the dose of epidural local anesthetic administered.

Methods: Sixty subjects receiving lumbar epidural bupivacaine were randomised to one of three groups: A: 10 ml 0.5% (50 mg); B: 10 ml 0.

Sum-frequency-generation spectroscopy of DNA films in air and aqueous environments.

Understanding the organization and orientation of surface-immobilized single stranded DNA (ssDNA) in aqueous environments is essential for optimizing and further developing the technology based on oligonucleotide binding. Here the authors demonstrate how sum-frequency-generation (SFG) spectroscopy can be used to compare the structure and orientation of model monolayers of ssDNA on gold in air, D(2)O, and phosphate buffered saline (PBS) solution. Films of adenine and thymine homo-oligonucleotides showed significant conformational changes in air versus aqueous environments in the CH stretching region.

Photoplethysmographic measurement of changes in total and pulsatile tissue blood volume, following sympathetic blockade.

Epidural anaesthesia, used for pain relief, is based on blocking the sensory and the sympathetic nerves in the lower part of the body. Since the sympathetic nervous system regulates blood vessel diameter, the sympathetic block is also associated with several haemodynamic changes. In the current study photoplethysmography (PPG) was measured on toes and fingers of patients undergoing epidural anaesthesia.

Increase in pulse transit time to the foot after epidural anaesthesia treatment.

Epidurally induced anaesthetic treatment is a routine treatment for pain relief during surgical procedure, based on blocking the sensory and sympathetic fibres that mediate pain. The epidural sympathetic block results in relaxation of the muscle walls in the lower limbs, which can be assessed by the resultant haemodynamic changes. In the current study, the difference tt,f in the transit time of the blood pressure pulses between the toe and the finger is measured by photoplethysmography (PPG).