Molecular genetics
The Laboratories are equipped with all the
resources necessary for cloning, subcloning, nucleic acid
isolation, blotting and mutagenesis, including polymerase
chain reaction. The equipment includes bacterial incubators,
cold room, dark room, electrophoresis supplies, high-speed
centrifuges, and commonly used radiotracers. DNA sequencing
is performed at the Sequencing Facility, part of the Columbia
Cancer Center.
Transgenic animals
In collaboration with the Columbia Transgenic
Facility in the adjacent Hammer Research Building, transgenic
mice are being produced as models of human disease.
Fluorescence in
situ hybridization (FISH)
The Laboratories host a dedicated microscope
and work area for this area of clinical and scientific investigation.
Currently, the resource is being utilized in the first step
of the genetic diagnosis of Glut-1 DS, since about 10% of
patients are FISH positive, eliminating the need for further
genetic testing.
Binding and tracer uptake assays
Protein binding and kinetic studies of transport
mediated by molecules of interest across cellular membranes
are used as a tool to investigate the function of patient
cells harboring mutations.
Tissue culture
Cells obtained from patients, mainly, but
not limited to, skin fibroblasts, are propagated and expanded
until functional or protein assays are performed. Gene
expression and amplification studies are currently being
carried out on these cells.
Cellular bank
All tissues with self-perpetuating capabilities
are susceptible to propagation and subsequent storage indefinitely
in deep freezers for future study. Devitalized patient
samples are also maintained under optimal preservation conditions.
Each sample is accompanied by an archived clinical description.
Protein chemistry
Resources include ultracentrifuges, a computerized
spectrophotometer, a disintegration counter, affinity purification
and blotting capabilities and enzyme activity assays, of
interest in certain metabolic abnormalities.
Voltage clamp
The laboratory includes a two-electrode voltage
clamp for use with injected Xenopus oocytes. RNA coding
for a wild type or mutated molecule of interest may be injected
into oocytes, normally devoid of such molecules and, after
a period of incubation, the functional properties can be
precisely studied. A computer using pCLAMP software controls
data generation and acquisition through a Warner amplifier.
The experimental setup is enclosed in an isolating cage
in which several experimental solutions are ready for application
to the oocyte. A temperature control system allows the
performance of experiments under the desired conditions.
Patch clamp
This laboratory is housed in an octagonal-shaped
glass enclosure for isolation and comfort. The vibration
table contains a Nikon TE-300 equipped with Hoffmann modulation
optics and epifluorescence filters. An Axopatch 200B amplifier
performs voltage control and current recording and a Pentium
IV microcomputer running pCLAMP 8 transforms the data for
analysis. The unit includes a Narishigue microforgue and
a Sutter processor-controlled micropipette puller. The
computer is linked to the Internet and to a networked high-capacity
drive for data backup.
Computing
The Neurology internal network links the Laboratories
to high-capacity backup servers and to the Internet via
optic fiber cable. The Laboratories include 13 PC and 2
Mac interlinked microcomputers dedicated to the various
scientific instruments or investigator desks. Most computers
contain Pentium III or IV processors and are equipped with
color printing and digital scientific presentation capabilities.
New patients can be videotaped and scientific photographs
stored on digital media. A public library of digital presentations
by members of the Laboratories on different topics of scientific
and clinical interest is being created.
Clinical databases
All patient-related information is maintained
in digital databases for study. Detailed records and results
of explorations with investigative value are classified
by disease and subdivided by key headings, in an effort
to facilitate research and to promote resource sharing.
Current databases include MELAS and other mitochondrial
disorders, Glut-1 DS, sickle cell disease and other poorly
understood conditions still being defined and characterized.
Clinical trials
All of the NIH-funded clinical trials are
coordinated to allow clinicians and scientists maximum interaction
with patients who chose to participate. Research patients
are brought to the Irving Clinical Research Center, a comfortable,
patient-oriented facility that includes a Pediatric branch
at Children’s Hospital of New York, and exposed to a variety
of diagnostic and quality of life assessments performed
by highly skilled investigators and staff. Over time, entire
families have elected to participate in the studies culminating
in the elucidation and treatment of numerous neurological
and other diseases.
Administration and management
The Laboratories could not function without
the assistance and care of dedicated staff members who look
after the documentation, transcription, patient contact
and numerous other invisible but essential tasks.