The SIU School of Medicine Research Imaging Facility (RIF) is a core
imaging and quantitative analysis facility that provides approximately 2900
square feet of laboratory space in the 825A building. The RIF is equipped with a
variety of microscopy and imaging systems including a Leica TCS SP5 spectral
laser scanning confocal microscope (LSCM), an Olympus Fluoview LSCM,
Hitachi scanning and transmission electron microscopes, an image analysis
workstation, a real-time PCR system and chemiluminescence, bioluminescence
and near-infrared imaging systems.
The facility is available to all trained faculty, staff and students through card
access control. Daily operations and maintenance of the facility are overseen by
a full-time RIF Supervisor, Craig Whitworth, who holds a Masters degree and
possesses 29 years of experience in basic research and microscopy. The RIF
Supervisor provides education and training for the equipment in the facility and is
also available for technical support, protocol development and contractual
research projects. Trained users have 24-hour access to the equipment, 7 days
a week. Scheduling of RIF facilities and equipment is accomplished using Oracle
on-line calendar software.
The equipment currently available in the RIF is as follows:
1. A Leica TCS SP5 spectral laser scanning confocal microscope that uses
acousto-optical components and a patented prism system in place of
excitation and emission filters, providing much higher transmittance efficiency
and far greater sensitivity than conventional confocal microscopes. Eight
laser lines (405, 458, 476, 488, 496, 514, 543 and 633nM) and 5 detection
channels accommodate a wide diversity of fluorophores and allow for the
simultaneous detection of up to 5 separate emission spectra. Equipped with
a DMI 6000 inverted microscope with a motorized stage, a temperature
control chamber and CO2 control system, this system is ideal for live imaging
experiments. A Windows-based format facilitates basic confocal applications
as well as advanced functions, such as FRET and FRAP.
Oracle Resource Name: RIF-Leica LSCM
2. A Hitachi S-3000 scanning electron microscope with secondary and
backscatter electron detectors. This scope is equipped with a large chamber
area, low-vacuum viewing mode and a cold stage option. The scope is,
therefore, capable of viewing standard samples (fixed, dehydrated and
coated) or “dirty” samples that have not been fully processed.
Oracle Resource Name: RIF-SEM
3. A Hitachi H-7000 transmission electron microscope, complete with scanning
TEM (STEM) and small sample SEM capabilities. The RIF maintains an
adjacent dark room for TEM negative and print processing.
Oracle Resource Name: RIF-TEM
4. An MCID image analysis workstation equipped with Nikon Microphot
compound microscope, Bencher M2 copy stand and Roper Scientific
CoolSNAP and CoolSNAPcf digital cameras. This system enables image
capturing and quantitative densitometry and morphometry analyses from a
variety of media, including glass slides, photographic prints and x-rays.
Oracle Resource Name: RIF-MCID
5. An Applied Biosystems StepOnePlus, real-time polymerase chain reaction
(rtPCR) system. This is a 96-well system that is capable of real-time or endpoint nucleic acid assays using FRET-labeled primers (such as Applied
Biosystems ‘Taqman’ products) or SYBR green.
This system can
accommodate up to 6 different annealing temperature zones on a single
plate.
Oracle Resource Name: RIF-Step One Plus
6. A Syngene G:Box iChemi XT chemiluminescence imaging system that is
equipped with a 4.2 megapixel, cooled (-28°C absolute and regulated) CCD
camera and 16 bit A/D, providing a contrast of over 65,000 grey levels. The
system is equipped for capturing and analyzing luminescent, fluorescent
(trans-illuminated UV or epi-illuminated blue, green, red and near-IR) or
bright-field images from a variety of materials, including Western blots, DNA
gels, culture dishes or multi-well plates. Users can employ relative intensity
values or calibrate their images for molecular weight and concentration
measurements.
Oracle Resource Name: RIF-Syngene G:Box chemi-imager
7. A Li-COR Odyssey, near-infrared imaging system. This system takes
advantage of near infrared fluorescence to obtain greater signal sensitivity by
imaging in a range that greatly reduces auto-fluorescence (700 and 800 nM).
The Odyssey can accommodate blots, gels, culture dishes, multi-well plates
and tissue sections. Double-labeled samples can be used and the high
signal-to-noise ratio allows images to be calibrated to the pico-gram range.
Oracle Resource Name: RIF-Odyssey
8. An IVIS Lumina in vivo bioluminescence imaging system (Caliper
LiveSciences, Hopkinton, MA). This system is specifically designed to noninvasively observe and quantify the progression of physiological events (such
as tumor growth, infection, nerve regeneration) at multiple time points within
the same subject. The system features a highly sensitive, cooled (-90°C)
CCD camera that can effectively detect low-level light emissions in small
rodents from luciferase-tagged cells after the administration of luciferin. This
system is also equipped with a xenon lamp and filter sets for GFP, DsRed,
CY5.5 and ICG for in-vivo fluorescence studies. The Lumina is housed in the
Laboratory Animal Medicine facililty.
Oracle Resource Name: RIF-Lumina
9. An Olympus Fuoview laser scanning confocal microscope (LSCM). This PCbased system is equipped with an inverted compound microscope, a singleline argon laser (488 nM), a dual line krypton laser (568 and 647 nM), 2
photomultiplier tube (PMT) detector channels and transmitted laser light
detection. Available emission filters (center/bandwidth) are 530/40 nM,
605/52 nM and 700/75 nM.
Oracle Resource Name: RIF-confocal
10. A Promega GloMax Multi Jr. fluorometer provides a fast and simple method
for determining the raw or quantified fluorescence from sample solutions.
The GloMax is equipped with multiple filter kits for measurement of UV, blue,
green or red fluorescence. Solutions can be measured in standard 10x10
mm methacrylate cuvettes or 12 mm test tubes. Smaller volumes (50-250ul)
can be measured with the available minicell Adapter Kit.
Oracle Resource Name: RIF-Promega GloMax fluorometer