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Redox & Bioenergetics Shared Resource and Free Radical Research Center


At the Redox & Bioenergetics Shared Resource

The RBSR offers state-of-the-art equipment, including a Seahorse Bioscience XF-96 analyzer and a Shimadzu UHPLC MS/MS detector.

The Seahorse XF-96 analyzer can detect pH changes and oxygen consumption rates for the mitochondrial analysis of eukaryotic cell lines, and the Shimadzu UHPLC MS/MS is used to develop and analyze drugs and compounds.

Seahorse XF-96 Analyzer

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Shimadzu UHPLC MS/MS Detector

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At the Free Radical Research Center

These instruments are available for use at the FRRC.
Agilent 1100 High Performance Liquid Chromatography
High-performance liquid chromatography (HPLC) is the technique of choice for the identification, quantification, and resolution of many small molecules involved in free radical research including probes for reactive oxygen and nitrogen species, as well as small proteins absorbing in visible light, including cytochrome c. Each of the two HPLC systems is equipped with a quaternary pump, a thermostatted well-plate autosampler, thermostated column compartment, diode array absorption detector and fluorescence detector. These HPLC systems are also used in the purification of newly synthesized compounds.
Agilent 8453 UV-Visible Spectrophotometer
This instrument is a single-beam PC-controlled, UV-visible spectrophotometer with collimating optics. It has diode-array technology which allows it to perform relatively fast kinetic measurements (full spectrum scan every 1 sec). This machine is equipped with a temperature controlled multicell (8 position) transport system, which can be used for enzyme kinetic studies. It has a wavelength range of 190-1100 nm and a spectral resolution of 1 nm.
Alpha Innotech Corp. Gel Documentation System AlphaImager
The gel documentation system includes a high performance CCD camera (zoom lens 12.5-75 mm with close-up +2 diopter lens), MultiImage® Light Cabinet and thermal printer for high quality printouts. The light cabinet allows for transillumination with UV and white light for fluorescence and colorimetric stains. It also has epi-illumination for analysis of blots, films or plates. The Alpha EaseTM software features a movie mode, on screen annotation, 1D-multilane analysis and spot densitometry. The imager is located in the Biophysics Department (2nd floor, MFRC).
Applied Photophysics Stopped-Flow Spectrophotometer
The Applied Photophysics, SX18.MV, is used for the kinetic analysis of fast reactions. In general, conventional UV-Vis spectrophotometry cannot measure the kinetics of reactions that occur in less than 10 seconds. This is because it takes at least 2 seconds to mix the components and place them in the instrument for analysis, referred to as the dead time). The stopped-flow, using a combination of high-pressure gas-driven syringes and mixing technology, has a dead time of less than 2 ms, and so is able to measure accurate kinetics of reactions that occur in as little as 10 ms. The instrument is able to work in both single and sequential mixing modes. The detection capabilities involve absorbance (photomultiplier or photodiode array) and fluorescence (cut-off filters or a second monochromator for choosing emission wavelength range) measurements.
Beckman Coulter DTX 880 Multimode Plate Reader
The plate reader is equipped with excitation and emission filters for the most commonly used fluorescent probes used in free radical research, and can operate in both absorption, fluorescence and luminescence modes. It has the capability of thermostating the plate during the measurement, as well as mixing the plate before reading. It can read fluorescence from both top and bottom of the plates. The instrument accepts most plate formats, including 6-, 24-, 96- and 384-well plates, with the additional option of measurement of 1536-well plate.
Bruker Elexsys Electron Paramagnetic Resonance System
This machine is a state-of-the-art EPR instrument that can be used to detect directly free radicals. The instrument is housed in the National EPR Center in the Biophysics Department (2nd floor, MFRC).
ESA HPLC System with Electrochemical CoulArray Detector
The HPLC system with electrochemical detector enables identification and quantification of electroactive compounds (antioxidants, amines, quinones etc.). The system consists of two pumps and high pressure mixer enabling gradient elution, thermostated autosampler, and an electrode array detector. The detector involves eight coulometric, working independently, porous graphite cells connected in a series. The system offers both high sensitivity and selectivity towards the redox active compounds. The system is also equipped with high voltage BDD amperometric electrode for measurements of the compounds characterized by high redox potential. Additionally, Hitachi UV-Vis absorption and fluorescence detectors with a post-column flow splitter allows for simultaneous detection of electrochemically active and UV-Vis absorbing and/or fluorescent compounds.
HyperChem 8.0 Software
The quantum chemistry calculation software package HyperChem 8.0 (Hypercube) enables prediction of the properties of molecules based on molecular mechanics, through semi-empirical to DFT and ab initio computational methods.
Nikon Fluorescence Microscope
The fluorescence microscope can be used for imaging the cells loaded with fluorogenic/fluorescent probes. It is equipped with numerous filters for most fluorophores in UV and visible range. Both brightfield and fluorescence pictures can be measured. Additionally, the system can monitor fluorescence resonance energy transfer (FRET) phenomena. The system is operated and data analyzed using MetaMorph software. The microscope is located in the darkroom in the Biophysics Department (2nd floor, MFRC).
Oxygen Meters
The oxygen electrodes enable the monitoring of changes in oxygen concentration in chemical systems (Rank Brothers LTD, reaction chamber volume: up to 1 ml) as well as in small volume biological samples, for example mitochondrial suspension (Mitocell MT-200, Strathkelvin Instruments, reaction chamber volume: 50 µl or 100 µl). Both systems are connected to and controlled by a computer.
PerkinElmer LS 55 Luminescence Spectrometer
This luminescence spectrometer is a very versatile instrument that allows users to measure fluorescence, phosphorescence, and chemi- or bio-luminescence. The instrument is based on monochromators, which enables collection of excitation and emission spectra and automatic building of fluorescence excitation-emission matrices (FEEM) spectra. An interchangeable well-plate accessory allows the user to measure samples in six 96 well-plates, cuvettes, TLC plates, electrophoresis gels, or any other flat sample type.
PerkinElmer Spectrum GX system FT-IR Spectrometer
FT-IR, or Fourier Transform Infrared, spectroscopy is a powerful technique for identifying the molecular composition of many organic and inorganic materials. Basically, the spectrometer measures frequencies at which the sample absorbs the radiation and the intensities of the absorptions. Determining these frequencies allows identification of the sample's chemical makeup, since chemical functional groups are known to absorb light at specific frequencies. The spectrometer is located in the Biophysics Department (2nd floor, MFRC).
PlasLabs Controlled Atmosphere Chamber
This chamber can be used for experiments under anaerobic (argon) conditions, or for manipulating hygroscopic compounds. Additionally, an analytical balance is kept inside in order to weigh air- and/or water-sensitive compounds.