Service Facilities for Biotechnology Research

Iowa State University

A major initiative of Iowa State University has been the establishment and support of state-of-the-art instrumentation for biotechnology research. The Iowa State University Instrumentation Facilities for Biotechnology Research are located on the university’s campus in Ames, Iowa. The facilities are open to faculty and students from the university, other educational institutions and industry scientists.

In addition to instrumentation facilities, the university has in place an organization whose primary function is the support of researchers’ equipment needs. The Research Equipment Assistance Program (REAP) maximizes use of research and teaching equipment by maintaining an equipment location database and loan program through which interdepartmental loaning and sharing of equipment is conducted. The REAP office has in inventory various pieces of research equipment that are available for loan.

The following are brief descriptions of the services offered by each of the instrumentation facilities. For additional information, see www.biotech.iastate.edu/service_facilities.

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Animal Gene Transfer Facility

The facility provides a variety of services to individuals who wish to utilize transgenic animals as part of their research program. The facility maintains equipment necessary for the creation of transgenic animals via microinjection or somatic cell nuclear transfer. The facility is available to life science researchers for technical service and for training of a limited number of individuals. All work to be performed in this facility must be scheduled with the professor-in-charge, as the facility is not staffed full-time.

Equipment housed in the facility includes two microinjection workstations, micropipette pullers, a microforge, stereo-zoom microscope and CO2 incubator. Additional supporting equipment located in nearby laboratories to which access may be granted includes a heated microcentrifuge, tri-gas incubators, stereo-zoom microscopes, inverted microscope, biosafety cabinet, osmometer, pH meter, embryo freezing machine, cell fusion machine, gel electrophoresis equipment and UV transilluminator.

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Atmospheric Air Quality Laboratory

The laboratory is situated at the National Swine Research and Information Center and serves as a focal point for collaborative research and training in the area of air quality engineering and livestock odor. The laboratory specializes in continuous air quality monitoring, environmental


analysis, quantification of organic compounds with gas chromatography and mass spectrometry, and simultaneous chemical and olfactometry analyses.

Field Air Quality Measurements

The laboratory is equipped with a variety of real-time analyzers for gaseous ammonia, hydrogen sulfide, carbon dioxide, methane, volatile organic compounds, particulate matter and ozone. It is also equipped to conduct field air sampling and analysis of organic gasses with sorbent tubes and SPME.

Multidimensional Gas Chromatograph-Mass Spectrometry-Olfactometry (MDGC-MS-O)

The MDGC-MS-O system includes custom-made GC-MS system with heat-cutting capability based on a Dean switch capable of simultaneous chemical and olfactometry analyses.

Solid Phase Microextraction (SPME)

SPME is used for extractions of gases emitted from a variety of samples, including livestock and poultry manure, air, food, packaging, bio-based products, live insects, plants, breath and general applications related to quality control and quality assurance.

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Atomic Force Microscopy Facility

Three atomic force microscopes are available for use in the Roy J. Carver Laboratory for Ultrahigh Resolution Biological Microscopy of the Institute for Combinatorial Discovery.

Digital Instruments DimensionTM 3000 Scanning Probe Microscope

The Dimension 3000 scanning probe microscope (SPM) brings together all SPM techniques in a single platform and handles a wide range of sample sizes and types. A rigid, low vibration construction of the Dimension 3000 SPM ensures the highest quality images and measurements.

Samples up to eight inches in diameter can be scanned in ambient air or fluids using the Dimension 3000 SPM. The Dimension 3000 SPM requires little or no sample preparation, and the simple vacuum mounting system allows easy and convenient setup. Superior linearity and resolution in all three dimensions are obtained, even for large samples. Integrated top-view video optics with motorized zoom and 1.5 µm optical resolution help identify areas of interest for detailed scanning quickly and easily. Changing scanning techniques, for example from AFM to STM, requires no tools.

The NanoScope IIIa system controller is a main part of the SPM system providing the software and electronics that drive the microscope. Digital tracking and feedback control ensure accuracy and speed at all scan sizes and positions on the sample.

Digital Instruments MultiMode with a Tapping ModeTM

The MultiMode system features multiple scanners that permit the user to tailor the system for individual research. Scanners with large scan ranges up to 120 microns on the X–Y axes, and a Z range up to 6 microns, as well as high-resolution scanners with 0.5 micron X–Y axes and submicron Z range are available. The vertical-engage “JV” and “EV” scanners allow the tip to be positioned at any point on the surface, without adjusting for lateral movement during approach. The MultiMode is controlled with a NanoScope IIIa controller. This controller provides 16-bit resolution on all three axes, with three independent 16-bit digital-to-analog converters (DACs) in X and Y for control of the scan pattern, scaling and offset. This configuration provides 16-bit resolution of the lateral scanning motion at any scan size.

Digital Instruments DimensionTM 3100 Scanning Probe Microscope

The Dimension 3100 is controlled with a Nanoscope IV controller. The NanoScope IV features up to ten-times-faster scanning, as well as increased functionality, bandwidth, flexibility and expandability.

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Center for Crops Utilization Research

The Center for Crops Utilization Research (CCUR) is a resource to assist ISU researchers and off-campus businesses in developing new value-added processes, products and markets for Midwest crops, especially corn and soybeans. CCUR strives to add value to grain and other crop-derived materials by conducting grant and contract research; offering short courses, workshops, seminars and training experiences; providing analytical, pilot plant processing, and consumer evaluation services; providing technical consulting services and information retrieval; and operating small-business incubator services. The center has grain, food and material processing equipment in state-of-the-art laboratory and pilot plant facilities.

The pilot plant facilities include a 5,000 square-foot wet-processing pilot plant (soy protein isolation, corn wet milling, brewing, etc.); a 2,600-square-foot dry-processing pilot plant (dry corn milling, drying, grinding, sieving, etc.); a 900-square-foot hazardous solvents extraction facility (vegetable oil extraction grain, plant material extractions); 3,000-square-foot product development laboratory (plastic extrusion, molding, and film blowing; building material processing; etc.); and various process development and analysis laboratories (chromatography, grain analysis, vegetable oil refining, baking, laboratory-scale process development, etc.). A small theater and conference facilities are available for technology transfer activities.

CCUR partly administers and works closely with the ISU Fermentation Facility, the Iowa Grain Quality Initiative and the Grain Quality Laboratory.

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Chemical Instrumentation Facility

The Chemical Instrumentation Facility has more than five million dollars worth of analytical instrumentation available to faculty, graduate students, industry and other educational institutions. The staff of five highly qualified professionals supports university research by keeping the analytical equipment available and operable and by providing application support and user training. Services available to the research community include the following:

Magnetic Resonance

Eight NMR spectrometers of varying frequencies from 60 to 600 MHz are available. Magnetic resonance spectroscopy allows the use of atomic nuclei as magnetic probes within a molecule. Chemical analysis and spatial orientation are determined by using this technique. A fully equipped EPR system also is available. Equipment located in the facility includes a Bruker Avance 600 (solids) NMR, a Bruker DRX-400 NMR, a Varian VXR-300 NMR, a Varian VXR-400 NMR, a Bruker AC-200 NMR, two Varian EM-360 NMRs and a Bruker ER-200 EPR. A Bruker Avance 700 MHz NMR is available in the Molecular Biology Building.

Mass Spectrometry

The mass spectrometry lab is equipped to provide both low- and high-resolution GC-MS on mixtures and high-resolution measurements for determining the elemental composition of pure samples. In addition, electrospray and APCI are used for the ionization of higher molecular weight compounds, particularly those of biological origin. Two dedicated LC-MS are now available for special projects. MS-MS (parent-daughter relationship) experiments are routine. A fully equipped MALDI-TOF for the analysis of biopolymers also is available. Equipment located in the facility includes a Finnigan TSQ-700 GC-LC-MS, a Kratos MS-50 MS, a Finnigan Magnum ITD GC-MS, a Micromass GCT-MS, a Shimadzu LCMS2010, a Finnegan LCQ LCMS and a Bruker PROFLEX-DE MALDI-TOF.

X-Ray Diffraction

A completely equipped X-ray diffraction laboratory provides instrumentation for the study of the molecular structures of small molecules and powders. The equipment includes a Bruker APEX II CCD single-crystal diffractometer equipped with low-temperature devices and a Scintag SDS-2000 powder diffractometer available for general use.

Spectrophotometry

A variety of spectrophotometers is available for routine use. These instruments provide “fingerprint” spectra for characterizing and identifying compounds. These instruments currently include a Bruker IFS 66V FT-IR, a Hewlett-Packard HP-8452 Diode Array UV-Vis and a Jasco J-710 circular dichroism spectrophotometer.

Elemental Analysis

A Perkin-Elmer Model 2400 Series II CHN/S elemental analyzer is available for sample submission or for investigator use. Normally, the instrument is configured for carbon, hydrogen and nitrogen, but sulfur also can be analyzed upon special request.

Computation

In addition to computer systems associated with the instrumentation, numerous PCs and workstations are available for network-based data processing and modeling. All data acquired in the facility are automatically backed up to tape or CD.

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Confocal Microscopy Facility

The Confocal Microscopy Facilities of the Office of Biotechnology, the Plant Sciences Institute and the Institute for Combinatorial Discovery have three confocal microscopes available for use by on- and off-campus researchers.

The confocal microscopes are located in 0117 Molecular Biology Building, the Roy J. Carver Co-Laboratory and the Roy J. Carver Laboratory for Ultrahigh Resolution Biological Microscopy. Researchers can choose which microscope best fits their research needs.

Confocal microscopes remove out-of-focus fluorescent light from the image, allowing clearer imaging of the sample, including the ability to view structures and components that were obscured by excess fluorescence that would be generated by standard fluorescence microscopes. Some applications of confocal microscopy include fluorescence microscopy when spatial distribution of cellular or other structures is important to the research being conducted, acquisition of a stack of images that can be used for 3D-reconstruction, viewing structures in thick tissue and removing background and cross-talk fluorescence or fluorescence from other structures that obscure viewing of the desired structure.

Leica SP5 X Confocal Microscope in the Molecular Biology Building

The confocal microscope in the Molecular Biology Building allows for real-time optical sectioning of fixed and living specimens, providing significant improvements in optical contrast and resolution over traditional light and fluorescence microscopy. The facility is equipped with a Leica SP5 X confocal microscope system with inverted microscope front end. New technology available on this system includes a white light laser. Researchers are no longer limited to three or four lasers at three or four fixed wavelengths but can tune the white light laser to any wavelength between 470 and 670 nm, which has a similar effect to having 200 lasers and 200 wavelengths. This allows researchers to select the excitation wavelength that is best suited to their samples and will result in maximum fluorescence emission. On the emission side, the system has an Acousto-Optical Beam Splitter (AOBS) which allows researchers to precisely set the emission wavelengths they would like to capture rather than be limited to preset emission ranges determined by fixed filter sets. Both of these technologies help to separate fluorescence signals with close excitation and/or emission spectra.

Other features include faster scanning, higher resolution and increased sensitivity. Researchers will now be able to work with live cells over an extended period using live cell equipment including a heated stage with microcontainment system, active gas regulation, cell cultivation chamber and microinjection system. For capturing images of fast moving samples or events that occur in a fraction of a second, the resonant scanner enables video rate scanning with speeds up to 16,000 lines per second. In addition, the system has FRET, FRAP, timelapse, hyperspectral signal separation software, 3D imaging, colocalization, deconvolution, region of interest (ROI) scanning, brightfield and DIC capabilities. A 405 laser provides UV excitation and an Argon laser provides CFP excitation and extra power for bleaching. The system also has an X-Y scanning stage and automatic composition software to capture high resolution images of samples that are too large to fit within one field of view and a color digital camera.

Nikon C1si Confocal Microscope in the Roy J. Carver Co-Laboratory

The facility at the Plant Sciences Institute is equipped with a Nikon C1si confocal microscope with both standard fluorescence confocal detection and spectral imaging capabilities. The 440, Argon, 561, and 638 lasers offer excitation wavelengths of 440, 457, 476, 488, 514, and 638 nm. Capture of transmitted light images is also available. The system offers 40x and 60x water dipping objectives in addition to 10x dry, 20x dry, 60x water, and 100x oil objectives. While in standard confocal detection mode, the system can capture up to three fluorescence channels and one transmitted light channel simultaneously. Additional capabilities include time lapse, FRET and FRAP. The system’s spectral imaging mode is useful for separating closely overlapping emission wavelengths from multiple fluorescent probes and/or autofluorescence. In spectral imaging mode, the system can simultaneously acquire up to 32 channels (2.5nm, 5nm, or 10nm individual channel widths) of fluorescence spectra in a single pass. Additional capabilities in spectral imaging mode include FRET and time lapse. A digital camera also is available on the system for standard (non-confocal) microscopy images.

Prairie Technologies Scanning Confocal Microscope in the Roy J. Carver Laboratory for Ultrahigh Resolution Biological Microscopy

A Prairie Technologies scanning confocal microscope is part of an optical workstation attached to an inverted Nikon Eclipse 200 microscope. Excitation wavelengths of 488, 568 and 633 nm are provided by Argon, Argon/Krypton and HeNe lasers. The instrument is completely computer controlled, including choice of dichroic mirrors, filters, pinhole size, scan size, integration time, photomultiplier voltage and z-focus. Images are stored on a hard drive and are available for export via Internet and CD. Image software is available that allows 3D reconstruction of confocal images generated with the confocal microscope.