Imaging and Microfluidic Core
The purpose of the Microfluidics and Imaging Core is to provide the personnel, skills, training, instrumentation and protocols to enable high-throughput optical analysis and experimentation on live cell samples, subjected to multiple perturbations across a variety of model and mammalian systems. This is to be implemented utilizing automated microscopy of samples mounted in multiwell slides and microtiter plates, and within microfluidic devices. In addition, instrumentation for cell sorting, cytometry, and laser-based cell manipulation also is managed within the Core.
Imaging Core. The focus of the Imaging Core has been the integration of the recently acquired hardware and software to enable high-throughput automated microscopy, and, with the Informatics Core, the development of an image acquisition pipeline for automated image data annotation, storage, and analysis. We have configured the imaging instrumentation to support the diversity of projects, cognizant of the need for different levels of resolution, growth criteria and depth of experimental throughput, as well as balancing the need for short- and longer-term experimentation.
The Core also operates a LEAP instrument (Cyntellect). This platform integrates three techniques essential to all cell-biological/biomedical research: imaging, transfection, and cell purification. These operations can be performed at high speed, with single cell precision, on both adherent and non-adherent cells in a standard microtiter plate format.
Microfluidics Core. Over the past year, all components of the Microfluidics Core have been brought into operation. A fully-equipped facility for the fabrication of multilayer polydimethylsiloxane (PDMS) devices has been established that houses a positive pressure hood to eliminate dust/particulates, a Thinky ARE-250 Mixer for PDMS pre-polymer preparation, a spin-coater, a dissection microscope for PDMS device layer alignment, a vacuum desiccator, an arbor punch to fabricate ports to connect to external control components, and a dedicated precision-controlled 80°C convection oven. The Microfluidics Core currently comprises four inverted microscopes (two Leica DMI-6000B, a Leica DMI-4000B, and a Zeiss Axiovert 135 TV) that have been fully equipped with all associated hardware components required to perform microfluidic experiments under full software control (LabVIEW, National Instruments).
The Center imaging and microfluidics tools have been integrated into workstations that support investigations using Halobacterium, yeast, and mammalian systems. The expanded capabilities support multiple users, and balances larger and smaller-scale needs. The Core’s enhanced training and standardized procedures have facilitated use of the Core’s capabilities. Core plans are to:
- Acquire a red laser to enable 4-color analysis on the BD FacsCalibur cytometer.
- Upgrade the Cytopeia Influx cell-sorter to digital electronics, and modify its light path to facilitate multicolor laser excitation.
- Acquire a BSL-2 hood and a CO2 cell culture incubator for safe and sterile cell manipulation and growth.
- Apply LEAP capabilities to address biological needs a variety of cell systems.
- Support a wide variety of cell manipulations in 96-well and 384-well microtiter plates and install a Matrix Hydra DT 96-channel fluidic.
- Devise how best to utilize scripts for customizing protocols for automated image acquisition using software.