LORRE

Capabilities and Facilities

The Laboratory of Renewable Resources Engineering (LORRE) has instruments, expertise, and protocols for addressing a range of research topics in bioprocess engineering, bionanotechnology, bioseparations, biorecovery, and bioenergy. In addition, the Laboratory, at Purdue University, has campus access to a range of instrumentation facilities through cooperative research projects or via shared facilities. LORRE's cooperation with Purdue's Discovery Park facilitates access to equipment which satisfies the analytical and imaging requirements of the research, as well as BL-2 areas for dealing with food pathogens, imaging facilities for monitoring and validating binding of labeled markers with microbial targets. These facilities and the laboratory environment of LORRE enable multidisciplinary research to be carried out in the context of graduate and undergraduate student education. The Laboratory is able to quickly start-up new projects, and address industrial research and development needs, as well as carry out fundamental mechanistic studies of biological systems.

Molecular Biology. LORRE is fully equipped for gene cloning and recombinant DNA manipulation of yeast and other microorganisms, including Bio-Rad Gene Pulser; Bio-Rad ChemiDoc Gel Documentation System with Quality One 1-D image analyses software; for performing PCR LORRE has BioRad iCycler (gradient temperature capable) and BioRad MyCycler thermal cycle; and electrophoresis for both DNA/RNA and protein. DNA Analysis: Purdue's Agricultural Genomics Center performs DNA sequencing, automated mapping, BAC fingerprinting (capable of up to 220K samples per year), plasmid DNA preparation (up to 200K samples per year), oligonucleotide preparation (capacity of 20K oligos per year), microarray slide preparation, and high density nylon filters.

Setting up an automated run on the fermentation system

Microbial Fermentors: LORRE has instrumented fermentors ranging from 1 to 40 L. The 40 L fermentor is a New Brunswick Scientific mobile pilot plant fermentor. The iL New Brunswick BioFlo 110 fermentors have BioCommand software for computer control and data logging. Shake Flask Incubators: Both temperature controlled floor and bench top units for shake flasks ranging from 250 mL to 2 L in size. Centrifuges: Beckman Types J2-21,Avanti J-30I, Avanti J-20XPI centrifuges, IEC HT (bench top) centrifuge and several Eppendorf bench top centrifuges. Autoclaves: SterileMax Tabletop 12 in dia x 18 in deep for sterilizing small items and Steris SG 120, 20 x 20 x 38 inch for sterilizing entire fermentation vessels.

The Bioflow 110 is a one liter bioreactor that can be used for fermentation and propagation of cells. We are equipped with three reactors each capable of monitoring and maintaining pH, dO2, agitation, temperature, and air flow rates

Pilot plant sized 30 liter reactor similar to the Bioflow 110. This reactor is also capable of monitoring and maintaining pH, dO2, agitation, temperature, and air flow rates 

Incubator Cabinets: For maintenance of Petri dish cultures. Low temperature incubator (-1.5C) also available. Sterile Transfer Hoods: For transfer of microbial cultures and preparation of seed cultures. BL-2 Safety Cabinet: Sterile hood to be used for working with pathogenic food organisms. Microscopes: Zeiss inverted stage; Olympia TO41 inverted stage light transmission microscope. Nikon Ty120 Florescence Microscope, up to 1000x magnification, with digital image capture. Freezers: SD Low Environmental Equipment Company Ultra-Low freezer (-70 C) and Fisher Isoterm freezer (-86 C) for microbial cultures and long term storage, and Revco (-30 C) freezer for plasmids; chest freezer for storing biomass samples. Freeze Driers/Concentrators: Lyophilizers for freeze drying samples and SpeedVac SPD2010 for vacuum drying/concentrating.

LORRE graduate student, Elizabeth Casey, working with the New Brunswick incubator

Other equipment that LORRE has access to includes: Plasmon Resonance: Biacore 2000 in Biology Department and IASYS SPR system in Food Science. Confocal Microscopy: Bio-Rad MRC 1024 UV/Vis System with Nikon Diaphot 300 inverted microscope, and Bio-Rad DVC 250 Viewscan Vis only with Nikon Diaphot 200 inverted microscope. Flow Cytometry: Coulter EPICS ELITE Cell Sorter, Bio-Rad BRYTE HS Flow Cytometer, and Coulter XL-MCL.

Fluorescence Inverted Stage Microscope

Florescence Microscopy: Advanced Nikon microscope with digital image captures and image analysis software is available through the Flow Cytometry Laboratories in the Hansen Building. This capability supplements fluorescence microscopes that are available in LORRE and which are used for preliminary observations. These include: an advanced Nikon (E600FN) fluorescence microscope with CCD camera (Coolsnap fx , Roper Scientific) for digital image captures, image analysis software (Metamorph, Universal Imaging Corp.), and a photomultiplier (PMT) tube (R3896), Hamamatsu) for photon counting applications. Electron Microscopy Facilities: Purdue's Biology Department has four transmission electron microscopes available for campus use. Two microscopes are used for high-resolution, cryo-electron microscopy of macromolecules such as viruses. These scopes (Philips CM200 and a Philips CM300) have field emission gun sources for maximizing high-resolution data collection as well as cryo-transfer specimen stages, anticontaminators and TV-rate and slow-scan CCD cameras. Two other microscopes, a Philips EM420 and a Philips EM410, are used for training and routine specimen examination. Access to this facility is available through X-ray Crystallography: Crystal growth facilities include three walk-in rooms with temperature-controlled environments at 20o, 12o and 4o C and an additional room for crystallization with cryoagents at -15o C. In-house X-ray diffraction data collection is done on three R-Axis image plate detectors mounted on 12kW Rigaku RU-200 rotating anode sources, each equipped with Yale focusing mirror optics and Oxford Cryosystem cryostream cooling devices. Purdue structural biologists take advantage of their close proximity to the high energy synchrotron source, Advanced Photon Source (APS), at Argonne National Laboratories, where data can be collected through the BioCars consortium on monochromatic, white radiation (Laue) and MAD beamlines.

Bioseparations. Preparative scale systems for open column and moderate pressure liquid chromatography; custom built/assembled preparative chromatography columns ranging from 500 ml to 40 L stationary phase volume; bench-scale membrane units, and gas phase adsorption systems. Cahn balance for measuring adsorbent characteristics. Analytical Liquid Chromatography: Systems for protein, peptide, carbohydrate, polysaccharide, organic acids and alcohols and other bioproduct separations. Detection based on refractive index and UV/Visible spectral absorbance. Most are equipped with autoinjectors and Waters Empower® software for HPLC control data acquisition, peak detection, and data analysis. Electrochromatography system. Barnsted e-pure system provides analytical grade water. Gas Chromatographs: Varian 3400 with TCD and FID detectors; Shimadzu GC-14A with FID and TCD detectors. Structure/ function studies are available through a proteomics core facility in the School of Agriculture that includes both LC mass spectroscopy and GC mass spectroscopy instruments.

Elizabeth Casey setting up parameters necessary to run low temperature water bath experiment

Engineering Computing Network. Provides computing capabilities for carrying out advanced modeling, and carrying out advanced analysis of engineering and biological data resulting from research in the laboratory. The Purdue Library system has on-line subscriptions to nearly all of the available on-line biology databases.

As is usual for such a laboratory, some of the equipment is dedicated for use solely for a specific research project at any one time. Other equipment is used for a few days and is required for multiple projects that utilize the same equipment or instrument, but require that advance scheduling be coordinated with the cognizant Laboratory Manager. In addition to laboratory facilities and hoods required in the normal course of research, there are equipment and instruments available for carrying out fermentation of microbial cultures, recovery and purification of products from such cultures, and analysis of materials generated by such fermentations.

Reactor Equipment. The Laboratory of Renewable Resources Engineering facilities at Purdue University include a scale-up facility with high-pressure reactors ranging from 5 mL to 20 L in volume with most pretreatment and enzyme mimetic work done in sandbath controlled 45 mL stainless steel tubular reactors. Instrument capabilities include liquid chromatographs (Waters, Ranin, and LDC, all with auto-injectors) for sugar, protein, and fermentation product analysis. Compositional analysis of the DDGS and other fibers is in a laboratory set up to follow the NREL standard laboratory procedures and has handled all previous samples on related projects. LORRE facilities include wet-bench facilities, molecular biology laboratories, fermentors up to 20 L, incubator-shakers, freezedriers, and cold sample storage. The Agricultural and Biological Engineering Department has a well-equipped machine shop capable of fabricating high-pressure fittings as needed for the research. The Engineering Computer Network of the Schools of Engineering at Purdue University provides the capabilities required for carrying out modeling and data analysis. High Pressure Reactors: 300 mL and 20 L Autoclave Company high pressure reactors; 2 L Parr high pressure reactor for carrying out processing of biomass materials. Steam Generators: Small and large generators capable of generating up to 50 psig and 300-psig steam, respectively. Size Attrition Equipment: Laboratory Wiley mill, large-scale refiner (a device that shreds cellulosic materials). Sand Baths: Tecaml laboratory scale (approximately 4 L volume) for carrying out high temperature reactions (up to 260 C). Padder: Atlas Laboratory wringer for derivatizing textile fabrics and fibers (threads). Furnace: Rapid temperature type high temperature furnace.

Shop. The Laboratory of Renewable Resources Engineering has access to shop capabilities of the Agricultural and Biological Engineering Department which can carry out fabrications of systems and schematic units having an approximate volume of 500 gallons. Equipment includes an arc welder (Miller Model SR200), and a TIG (tungstun inert gas) welder that allows welding of aluminum and stainless steel. The TIG unit is a Miller Synchrotron 300. The shop is fully capable of welding different types of stainless steels, and vessels as large as 500 gallons can be handled.

The shop has 2 metal lathes with a bed capacity of 14 inches (turning capacity) and a bed length of 36 inches. In addition, the shop has access to a milling machine, which is a Bridgeport with a 31-inch horizontal dimension. Horizontal and vertical bandsaws are also available including a Johnson Model J, and a vertical bandsaw (dual, Model 26). The shop also has access to a radial drill press (Fosdick, 3-foot) and a 48-inch shear. These capabilities are further complemented by the Central Machine Shop, which is a full service machine shop, with an expert design and drafting staff, and includes virtual development services to test feasibility of conceptual ideas.

Return to Top of Page