Neurobiology Imaging and Tissue Processing Core (UAB-MRRC Core C)
Co-Directors: Lucas Pozzo-Miller, PhD and Harald Sontheimer, PhD
Department/Center Association: Mental Retardation Research Center (MRRC)
Established: 2000
Mission
The objective of the UAB Neurobiology Imaging and Tissue Processing Core is to
provide state-of-the-art equipment and technical support for several experimental projects
across campus related to mental retardation and the UAB Mental Retardation Research Clinic
Community mission statement. The Core does not charge fees for instrument usage or
technical consultations; however, users provide the necessary reagents for tissue processing.
Facility Description
The Neurobiology Imaging and Tissue Processing Core is housed in two facilities in the
Shelby Interdisciplinary Biomedical Research Building, within laboratory space allocated and
maintained by the Department of Neurobiology. The first floor facility (SHEL-130) houses all
resources related to tissue processing, the Leica digital fluorescence microscope, one of the
laser scanning confocal microscopes, and the supporting equipment for preparation of ultra-thin
sections for transmission electron microscopy. The tenth floor facility (SHEL-1062) houses the
multiphoton imaging/electrophysiology microscope along with all its supporting equipment.
Finally, the transmission electron microscope is located in the basement of the Civitan
International Research Center (CIRC-B16). Additional instruments available to the UAB-MRRC
community are located in neighboring buildings, as part of the UAB High Resolution Imaging
Facility. The MRRC Director and several of the MRRC Investigators serve on the Advisory
Group and User’s Group of the UAB High Resolution Imaging Facility, respectively.
The UAB Neurobiology Imaging and Tissue Processing Core has the following imaging
instrumentation:
 A tissue processing laboratory equipped with perfusion set-up for small rodents (rats and
mice), sectioning areas, chemical bench, balances, centrifuges, embedding vacuum
ovens, compound light microscope, and staining set-ups (SHEL-130).
 A Pelco 3450 Microwave Processor for enhanced aldehyde fixation using microwaves,
equipped with feedback-liquid-cooling temperature control during fixation, staining,
dehydration, and pre-embedding for LM and TEM (SHEL-130).
 A Leica Jung CM3000 cryostat for obtaining frozen semi-thin sections (SHEL-130).
 An ultramicrotomy room with Reichert Ultracut S ultramicrotome equipped with a cryobox
for obtaining ultra-thin sections of plastic embedded tissues, as well as ultra-thin
cryosections of rapidly frozen material (SHEL-130).
 A custom-modified LifeCell CF-100 rapid freezing machine for slam freezing of cell and
tissues (SHEL-130).
 A Jeol 100CX transmission electron microscope (CIRC-B16).
 A fully equipped darkroom for developing, enlarging, and printing light and electron
microscope negatives (CIRC-B16).
 A Leica DMRB upright fluorescence microscope with dry and oil-immersion objectives
and equipped with a Optronics DEI-750 color CCD camera, frame grabber, Dell personal
computer, and Kodak DS 8650 PS color printer for digital image acquisition, analysis,
and printing (SHEL-130).
 A Lumistar BMG 96 well automated luminescent plate reader (CIRC 4th floor, Sontheimer
lab).
 An Olympus FluoView 300 Laser Scanning Confocal Microscope System mounted on an
Olympus BX50 upright fluorescence microscope with dry and oil-immersion lenses, and
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Pentium workstation with image analysis hardware and software. The system has Argon
(488nm) laser and Krypton (568nm) lasers for excitation. This is a fixed-tissue confocal
microscope, which belongs to the UAB High Resolution Imaging Facility (SHEL-130).
An Olympus FluoView 300 Confocal Scanning System mounted on a Olympus BX50WI
fixed stage upright fluorescence microscope equipped with water immersion lenses and
Nomarski DIC-IR optics; Chameleon tunable infrared laser for multi-photon excitation
microscopy (~720-950nm, 10W solid-state diode laser pumping a 90Mhz regenerative
mode-locked Ti:Sapphire laser, <80fsec pulse width, Coherent); a Hamamatsu Orca
enhanced infrared-sensitive, cooled CCD digital camera for visually guided patchclamping and low light, supra-video rate fluorescent imaging; an elevated platform
mounted on X-Y mechanical translators replacing the microscope stage; a temperaturecontrolled, perfusion brain slice chamber and peristaltic pump for artificial cerebrospinal
fluid perfusion; two Burleigh patch-clamp solid-state piezoelectric manipulators mounted
on the platform; an Axon Instruments Axopatch 1D patch-clamp amplifier; two
extracellular isolated stimulators; one dual-channel digital storage oscilloscope; one
analog-to-digital/digital-to-analog computer board; one Pentium workstation running
Windows XP for acquisition, analysis, and storage of laser scanning imaging data; one
G4 Mac running OSX for acquisition, analysis, and storage of electrophysiology and
imaging data from the cooled CCD camera; and a TMC vibration isolation table (SHEL1062).
A Leica vibrating blade microtome VT1000S for live tissue sectioning (SHEL-1062).
A Leica Laser Scanning Confocal Imaging Spectrophotometer TCS SP unit mounted on
an inverted microscope, and equipped with Krypton, Argon, and Helium/Neon lasers for
confocal microscopy. This is a fixed-tissue microscope, which belongs to the UAB High
Resolution Imaging Facility (SHEL-130).
A Leica Confocal Imaging Spectrophotometer TCS SP UV unit mounted on an inverted
microscope, and equipped with Krypton, Argon, and Helium/Neon lasers and a Coherent
Laser Group Enterprise UV laser for use with UV excitable dyes such as indo-1 and
DAPI. This is a fixed-tissue confocal microscope, which belongs to the UAB High
Resolution Imaging Facility (VA hospital).
Research Information
The Neurobiology Imaging and Tissue Processing Core is a major resource for the UAB
MRRC faculty. It provides state-of-the-art equipment unlikely to be acquired by any single
investigator's research program, as well as highly trained staff for its maintenance. The shared
use of the Core provides synergistic support to several projects within the MRRC community
including: advances and improvements of common methodological and conceptual strategies,
access to regular and reliable analytical methods for live cell imaging, quantitative image
processing, and additional collaborations between the individual projects.
The Core also provides simultaneous whole-cell recording and imaging of intracellular
ion concentrations, presynaptic vesicle release and recycling, or morphological changes from
single cells in culture or brain slices at sub-micrometer spatial and millisecond temporal
resolution. Investigators can do high-resolution laser scanning and live imaging for long-term
morphological studies of synapse formation, as well as time-lapse developmental studies of
dendritic spines, dendritic arborizations, axonal projections, or astrocytic processes.
Services and Fees
The Core will provide training and assistance to UAB MRRC investigators and their lab
personnel. The assistance will be offered on an as-needed basis and investigators are allowed
to run the microscopes by themselves after training. A sign-up scheduling system is available in
the Neurobiology server (contact Lucas Pozzo-Miller for information). There are no service fees
for the use of the Core. Interested investigators wishing to become MRRC investigators to gain
access to all its Cores (http://www.mrrc.uab.edu/cores.html) should send their request to the
MRRC Directors, Drs. Alan Percy (934-8900, apercy@uab.edu) or Harald Sontheimer (9755805, hws@uab.edu).
Contact Information
Co-Director: Lucas Pozzo-Miller, PhD
Email: lucaspm@uab.edu
Phone: 205-975-4659
Co-Director: Harald Sontheimer, PhD
Email: hws@uab.edu
Phone: 205-975-5805
Core Staff: Edward Phillips
Email: Phillips@nrc.uab.edu
Phone: 205-934-7933
Web Site: http://www.mrrc.uab.edu/corec.htm
Approved by: Lucas Pozzo-Miller, Core Co-Director
Date: February 7, 2007
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