Research Facilities, Technology and Equipment
A wide variety of behavioral and physiological data can be gathered using the resources available at David King Hall and the Krasnow Institute. Listed here are some of the resources commonly used by students in their research.
Neurolucida
We have the latest version of Neurolucida. Neurolucida's capabilities allow for reconstruction of entire neurons in 3-D. Additionally, as the neuron is traced, a battery of measurements are made automatically, so that data on morphometric parameters such as neurite length, diameter, and spine density are immediately available. The system comprises the Neurolucida software, a BX51 Olympus microscope, an automated stage, and a 22 inch LCD monitor used to display strikingly clear digital pictures. These system capabilities allow for unparalleled precision in quantification of neuronal morphology.
Skinner Box System
We have eight operant Skinner boxes used for self-administration experiments in rats. They are fully automated and can be used for cued and contextual conditioning with food or drug.
Fear Conditioning Systems
We currently have two working fear conditioning systems. We have two chambers operating with the San Diego system and two chambers using the recently acquired CleverSys system. Both systems are fully automated for computerized measurement of freezing behavior and behavioral analysis of analgesics and anxiolytics, and are suitable for use with both rats and mice. Here, a Long-Evans rat is shown in one of the San Diego chambers.
Morris Water Maze
Our Morris water maze system works with the HVS 2020 system to collect behavioral data for both rats and mice. We have four pools measuring 6 ft., 5 ft., 4 ft., and 3 ft. in diameter, with a depth of 2 ft. The pools can be painted white or black to track dark or albino animals. We have platforms for Atlantis and moving platform paradigms. A camera is mounted above the maze and tracks movements through the HVS imaging system. The pool is surrounded by large black spatial cues mounted on a white background. The HVS 2020 system can also be used for other tracking tasks such as analysis of open field behavior.
Open Field and Place Preference
We have an open field system which may be used to study anxiety levels in rats and mice, especially in response to treatment with a drug. Smith lab is currently using a conditioned place preference (CPP) system to study Pavlovian conditioning cues in relation to drug-seeking behavior in rats. CPP is a potentially useful test for modeling human drug-seeking, craving, and relapse, as drug-associated cues can maintain drug-taking behavior long after the absence of a drug.
Novel Object Response System
We have a novel object response system that has been used for rats and is undergoing pilot testing with mice. Animal behaivor is tracked by camera and analysed using the Clever Sys ObjectScan software.
Cryostat and Vibrotome
We have a two cryostats available for tissue sectioning. The newest model is a Tissue-Tek Cryo3 microtome/cryostat manufactured by Sakura, shown here. We also have a Vibrotome, which is ideal for sectioning fixed or perfused tissue such as Golgi-Cox stained brains.
Rat Colonies
Our animal housing facilities provide living space for several inbred rat strains, primarily Sprague-Dawley and Long-Evans. Facilities exist in both David King Hall and the Krasnow Institute, so experiments may be conducted at both locations. We have capacity for both single and group housing, as well as enriched environment housing. Breeding and animal care is conducted in-house, allowing for careful control of breeding conditions and convenient access to all rodent models for experimentation. A separate breeding room within the colony is maintained to reduce stress on breeding mothers and unweaned pups.
Transgenic Mouse Colonies
The rat colony at David King Hall also provides space for transgenic mouse colonies, maintained by Drs. Flinn and Fryxell. Room 2037 houses the Flinn lab mouse colony, consisting of several cohorts of TgAPP2576 mice, which produce amyloid plaques mimicking those found in Alzheimer's disease. Animals may be single or group-housed, and have "enriched" environments with igloos, wheels, and rubber bones in their cages. Transgenic mice may be purchased or bred in-house.
Genotyping
Trasgenic mice bred in-house can be genotyped in-house at the Fairfax or the Prince William campus. Tissue samples can be collected and DNA extracted very easily in Fairfax; in cooperation with the biology department on the third floor of David King Hall, DNA can be amplified using PCR and gels can be run in a few hours. Real-time PCR can be run at the Prince William campus.
Synchrotron Imaging
Dr. Flinn's trace metal analysis lab is devoted to the study of metals in learning and memory. To this end, students make frequent trips to the U.S. Department of Energy's Brookhaven National Laboratory in New York. There, they use both X-ray fluorescence and infrared spectroscopy to collect data on mouse, rat, and human tissue. Microprobe synchrotron X-ray fluorescence provides a way to quantitate levels of metals in tissue, while infrared spectroscopy allows us to measure levels of amyloid protein in different conformations.
Histochemical Analyses
We have a chemistry lab in David King Hall, along with resources in Krasnow, to do many kinds of histochemical analysis. Rodent neural tissue can be perfused or fresh-frozen, sectioned with cryostat or vibrotome, and stained with a variety of stains. Prepared slides can be imaged with any of several microscopes, an we can do bright-field, dark-field, and fluorescent microscopy. We are also setting up a darkroom that will allow us to process films from autoradiography in situ hybridization in David King Hall.
On the left you see neurons stained with Golgi-Cox solution, which can be traced and reconstructed with the Neurolucida system. On the right is an amyloid plaque stained with Congo red solution, which produces apple-green birefringence when viewed under polarised light.
