Equipment and Instrumentation
A variety of automated tools and equipment are employed in the high-throughput screening workflows. This includes a number of commercially available and custom-built instrumentation.
The Tecan EVO liquid handling system is utilized to automate the various protocols and procedures of the high-throughput biological assays. It contains a liquid handling arm (LiHa) that can work with both tubes and plates enabling rapid tube to multi-well plate sample transfers and dilutions. The LiHa is equipped with eight tips configured with a wide combination of fixed and disposable options. Bulk reagent additions can be performed with these devices or with integrated reagent dispensers. This system is also configured with a Robotic manipulator (RoMa) arm, which allows the integration of peripherals around all four sides of the Freedom EVO deck. An incubator is mounted at the end of the deck and maintains precise control over temperature (–20 to 95°C), CO2, humidity and shaking. The deck has been modified (right) by CNSE engineers to appropriately handle 24-well plates for reagent additions and extractions. A Tecan Safire2 is mounted on the rear of the deck and is utilized for fluorescence, luminescence and absorbance measurements.
The Olympus microscope system has the ability to image in reflected light from 50x to 1000x magnification and has fluorescence, bright field, dark field, and DIC capabilities. Imaging for these techniques is carried out on two different cameras including a 12.5 million pixel DP70 digital camera and a color digital camera. The system also has wide field as well as a disk scan confocal capability for 3D imaging. The confocal imaging is carried out on a cooled black and white Hamamatsu camera. The confocal analysis can be carried out at very high magnification/resolution or low magnification for multi-well plate applications. The system is also equipped with a motorized stage that can be controlled through the software as well as joystick control.
The Mx3000P RT-PCR system has a scanning PMT (350-700 nm) and a thermal cycler with a temperature range of 25-95°C at ± 0.25°C variation in sample temperature across a 96-well plate. The Mx3000P is specifically designed to ensure optimal results with multiplex assays using up to four targets in the same sample. In addition, the Mx3000P achieves linearity of amplification over seven orders of magnitude and possess the ability to detect two-fold differences in initial template quantity with 99.7% confidence.
The BMRL utilizes two multi-well plate reading systems for carrying out a variety of spectrophotometric assays based on fluorescence, luminescence and absorbance measurements. The first system is a TECAN Safire2 (left picture) which is a modular microplate detection system based on a quad-4 monochromator technology that offers a range of high speed detection techniques. The Safire2 is mounted on the deck of our Tecan EVO liquid handling robot to enable automated plate measurements. The second system is the Synergy HT from BioTek (right picture) and is based on a dual-optics design that allows for three different reading modes. This system also has a low noise PMT for high performance luminescence measurements.
This electroporation system is designed to deliver precise field strengths and pulse lengths required for the transfection of mammalian cells and transformation of bacterial and yeast cells. In the High Voltage mode, this system is optimized to electroporate E. coli and other bacteria and yeast.
This microscope is outfitted with phase contrast and brightfield capability and includes a unique swing-out condenser and long working distance that can accommodate a variety of vessels. The microscope contains a trinocular head for image documentation via camera attachments. In addition, the microscope includes six long working distance objectives: three brightfield planachromatic (10x, 25x, and 40x magnification) and three phase contrast planachromatic (10x, 25x, and 40x magnification). Microscope also features a quintuple-position turret for precise, single knob focusing and an inviting field of view. Illumination is provided by 6V, 20W variable intensity quartz halogen light with iris diaphragm and focusable collector lens.
A Minifors Fermentation System was purchased from ATR Biotech and is utilized for batch, fed batch and continuous culture of microorganisms. The system is composed of a 2.5l vessel, four independent peristaltic pumps, and an integrated controller for continuous monitoring and control of stirrer speed, temperature, pH, pO2, antifoam level and gas mixing.
An automated aquarium system was purchased from Marine Biotech and is utilized to maintain marine organisms year round at NDSU. Artificial sea water (ASW) is supplied to 16 aquarium tanks from a 200 gallon reservoir. The system recirculates the ASW through activated carbon and polishing filters to remove organics and particulates, respectively. Temperature, pH and salinity of the ASW are continuously monitored and maintained by the system. Approximately 300 coating array panels can be housed in the system simultaneously.
An automated water-jet apparatus was designed and fabricated at NDSU for screening microbial adhesion. The system consists of a robotic arm that is mounted on a stainless steel deck which can access coating array plates from one of three plate stacking hotels. The robotic arm inverts each well of an array plate over an offset, rotating nozzle that delivers a jet of water perpendicular to the coating surface. The duration and pressure of the water-jet are precisely controlled through a customized software program. Coating array plates are subjected to water-jet treatments after bacterial or algal incubation to facilitate removal of adherent biofilms.
A semi-automated barnacle push-off apparatus was designed and fabricated at NDSU. A hand held digital force gauge is mounted on the deck of a motorized stage. A built-in motorized actuator facilitates the application of a straight line shear force at a constant speed. The height of the stage can be adjusted to accommodate variations in coating thickness. The digital force gauge is interfaced to a CPU to record the peak force required to remove reattached barnacles from the coating surfaces.