Short History
This core facility was founded by a group of scientists in the Molecular Microbiology & Immunology department to facilitate their need for faster turnaround with primers, DNA sequencing, and fluorescent imaging. In a few years, the need for these services by the whole campus became apparent and a full-time core facility director was hired in 1995.
During the last few years there has been a substantial increase in use of this shared resource reflecting the quality, convenience and value of the services provided. We have carved out a niche for these services in the university by combining highest quality reagents and efficiency with expert assistance. We provide help with experimental design, convenient ordering methods, and fast turnaround times.
In addition to our fee-for-service work, we are glad to help with experimental design, data analysis, and application troubleshooting. Click on any of the "site menu" items on the right for more information about the specific services.
Staff
| Director, Thomas J. Keller, PhD.Joined the MMI Core Facility as Director in 1995 after running the DNA Services core at the University of Wisconsin, Madison for five years. Received his PhD in Medicinal Chemistry in 1985 from University of California, San Francisco. |
| Microscopist, Aurelie SnyderBecame the Microscopist for the Core Facility in 1998 after many years in Dr. Sample's group at the Casey Eye Institute. Aurelie has 30-some years of experience in microscopy in all areas, from clinical histology to electron microscopy, with the last 10 years focused on immunofluorescense and live cell imaging. She can advise you on your experiment design and protocols. She is a regular group leader at Jim Pawley's yearly Live Cell Microscopy summer course at UBC, Vancouver. |
| Core Lab RA, Tracy McFarlaneJoined the Core Facility in 2003 after a stint doing molecular biology in the School of Dentistry. She graduated from PSU with a BS in Biology in spring of 2003. |
| Core Lab RA, Brian AevermannJoined the Core Facility in 2007 after receiving a Master's Degree in Bio-informatics and Molecular Biology with Dr. E. Waters at the San Diego State University where he focused on comparative genome analysis methods. |
Instrumentation
DNA Sequencing
We acquired our DNA analyzer, an ABI 3130xl, in 2005. It is an 16 capillary system capable of running about 300 samples per day with read lengths of over 800 bases. It can also do multiplexed, fragment size (AFLP) analysis for high-throughput genotyping.
DNA & RNA Synthesis
We have two 4-column ABI 394 solid-phase DNA synthesizers. They each have 4 additional reagent ports for incorporating multiple labels, 5'-phosphate, biotin, and numerous base analogs. It is also able to synthesize RNA and mixed RNA and DNA. We are a very cost effective source for synthetic siRNA molecules.
Confocal Microscopy
We perform confocal microscopy on the Oregon Hearing Research Center's new OHRC Olympus confocal which is the Fluoview-1000 (FV1000) on an IX-81 microscope base; an electronic turret with 10x and 20x air and 40x and 60x oil objectives. It has 405, 440, 488, 515 and 559 nm excitation laser lines and can simultaneously image while initiating a laser event such as uncaging, FRAP, FLIP, and photoactivation. It can acquire a differential interference contrast (DIC) and overlay it with the fluorescence images.
Image Restoration (Deconvolution) Microscopy
The Core DV system is a high resolution wide field microscope for acquiring images of live or fixed cells and tissues. The basic system is based on the Olympus IX71 microscope with differential interference contrast (DIC) transmitted light and 3 lasers (402nm, 488nm, 532nm) for photoactivation (PA), photobleaching (FRAP) and total internal reflection (TIRF). The light source is a short arc Xenon lamp and the excitation filter wheel has 10 places while the emission has 6. Both wheels are exchangeable for fluorescent protein-specific or user-specific filters. More info about standard and live cell filter sets here. The camera is a Nikon Coolpix HQ for fast, sensitive acquisition. The patented stage is controlled by extremely accurate XYZ nanomotors for exact z-stack and point-visiting functions.
The stage area is enclosed by a Weatherstation™ environment cabinet providing controlled heat, moisture and carbon dioxide to facilitate live cell imaging over very short ( a few minutes) to very long (days) times. Deconvolution, using an algorithm based on Sadat and Agard's work, provides image restoration processes to the appropriate data sets. Additional software (Imaris™ from Bitplane) is available for in depth analysis of the images for many variables such as intensity, direction and speed vectors, structure volume and colocalization.
Within one year the core facility will add fluorescent lifetime imaging (FLIM) to the abilities of this system. The image processing can be done on either the core computer or an additional stand alone. Up to three users can be accessing the image data bank at one time initially and more licenses can be purchased to facilitate user access to their data.
Here is a qt movie of our the last Core Lab party