ABSTRACT
The tedious sample preparation for flow cytometry limits the throughput and thus its usage as a primary screening method despite its sensitivity and accuracy. With the growing focus on utilizing antibodies as a therapeutic modality in drug discovery, it is critical to develop a high-throughput flow cytometry (HTFC) workflow to cope with the increasing need to support antibody discovery programs. We have developed a seamless HTFC sample preparation and readout workflow using the HighRes modular robotic system and the IntelliCyt iQue Screener PLUS. To fully utilize the advantages offered by flow cytometry, we typically multiplex multiple cell lines of interest in one well to simultaneously quantitate on-target activity and nonspecific activity along with measurement of antibody concentration. The ability to measure multiple parameters coupled with speed and increased accuracy provides gains in productivity and helps speed up antibody lead discovery.
Subject(s)
Antibodies, Monoclonal/pharmacology , Drug Discovery , Flow Cytometry , Animals , Automation , Drug Discovery/methods , Drug Evaluation, Preclinical , Flow Cytometry/methods , High-Throughput Screening Assays/methods , Humans , Hybridomas , Immunoglobulin G/pharmacology , Mice , WorkflowABSTRACT
The application of 3D printing and microcontrollers allows users to rapidly engineer novel hardware solutions useful in a laboratory environment. 3D printing is transformative as it enables the rapid fabrication of adapters, housings, jigs, and small structural elements. Microcontrollers allow for the creation of simple, inexpensive machines that receive input from one or more sensors to trigger a mechanical or electrical output. Bringing these technologies together, we have developed custom solutions that improve capabilities and reduce costs, errors, and human intervention. In this article, we describe three devices: JetLid, TipWaster, and Remote Monitoring Device (REMIND). JetLid employs a microcontroller and presence sensor to trigger a high-speed fan that reliably de-lids microtiter plates on a high-throughput screening system. TipWaster uses a presence sensor to activate an active tip waste chute when tips are ejected from a pipetting head. REMIND is a wireless, networked lab monitoring device. In its current implementation, it monitors the liquid level of waste collection vessels or bulk liquid reagent containers. The modularity of this device makes adaptation to other sensors (temperature, humidity, light/darkness, movement, etc.) relatively simple. These three devices illustrate how 3D printing and microcontrollers have enabled the process of rapidly turning ideas into useful devices.
Subject(s)
Engineering/methods , Environmental Monitoring/instrumentation , High-Throughput Screening Assays/instrumentation , Printing, Three-DimensionalABSTRACT
Working memory ability matures after puberty, in parallel with structural changes in the prefrontal cortex, but little is known about how changes in prefrontal neuronal activity mediate this cognitive improvement in primates. To address this issue, we compare behavioural performance and neurophysiological activity in monkeys as they transitioned from puberty into adulthood. Here we report that monkeys perform working memory tasks reliably during puberty and show modest improvement in adulthood. The adult prefrontal cortex is characterized by increased activity during the delay period of the task but no change in the representation of stimuli. Activity evoked by distracting stimuli also decreases in the adult prefrontal cortex. The increase in delay period activity relative to the baseline activity of prefrontal neurons is the best correlate of maturation and is not merely a consequence of improved performance. Our results reveal neural correlates of the working memory improvement typical of primate adolescence.
Subject(s)
Memory, Short-Term/physiology , Neurons/physiology , Prefrontal Cortex/physiology , Sexual Maturation/physiology , Age Factors , Animals , Cues , Macaca mulatta , Male , Prefrontal Cortex/cytology , Psychomotor Performance/physiology , Visual Perception/physiologyABSTRACT
Executive functions including behavioral response inhibition mature after puberty, in tandem with structural changes in the prefrontal cortex. Little is known about how activity of prefrontal neurons relates to this profound cognitive development. To examine this, we tracked neuronal responses of the prefrontal cortex in monkeys as they transitioned from puberty into adulthood and compared activity at different developmental stages. Performance of the antisaccade task greatly improved in this period. Among neural mechanisms that could facilitate it, reduction of stimulus-driven activity, increased saccadic activity, or enhanced representation of the opposing goal location, only the latter was evident in adulthood. Greatly accentuated in adults, this neural correlate of vector inversion may be a prerequisite to the formation of a motor plan to look away from the stimulus. Our results suggest that the prefrontal mechanisms that underlie mature performance on the antisaccade task are more strongly associated with forming an alternative plan of action than with suppressing the neural impact of the prepotent stimulus.
