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.
Tecan Freedom EVO 200 Liquid Handling System
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.
Olympus Disk Scan Confocal Fluorescence Microscope
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.
Multi-Well Plate Readers
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.
BTX ECM 399 Electroporation System
VistaVision Inverted Microscope
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.
Marine Biotech Aquarium System
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.
Automated Water Jet Apparatus
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.
Barnacle Push-Off Apparatus
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.
Mussel Pull-Off Apparatus
A fully automated mussel pull-off apparatus was designed and fabricated at NDSU. To carry out adhesion measurements using this tool, the rod of each attached mussel is secured to an individual 5N load cell of a custom-built, tensile force gauge outfitted with six load cells to enable simultaneous measurements of all attached mussels to be made. The total force required to detach the byssus threads for each mussel is recorded (1 mm/sec pull rate) and the average pull-off force value (Newtons) for all attached mussels are calculated for each coating surface.