Biomechanical and Biochemical
- Biomechanical analysis
Madihally is the primary custodian and operator of an Instron 5542 mechanical testing equipment with a 100 N load cell. It is operated through Merlin 7.0 software, loaded on a dedicated computer. There is also a strain gage extensometer to perform creep analysis in addition to stress-relaxation equipments. There are different grips such as pneumatic grips, compression grips, and screw grips. There is also a custom built environmental chamber which allows testing to be performed at physiological conditions, necessary for testing various biomaterials.
With the help of OSU Center for Innovation and Economic Development, Inc, another Instron with variable load cell was purchased. Madihally was a co-PI on the proposal. This is operated as a core facility and can be used for impact testing for materials research. Through collaborative research grants, Madihally’s laboratory routinely has access to controlled stress-strain Bohlin Rheometer (Model C-VOR) with extended temperature option and TA Dynamic Mechanical Thermal Analyzer (Model DMA Q800 V20.20). Both instruments are extensively utilized to characterize hydrogel formulations.
- Biochemical analysis
Madihally is the primary custodian and operator of an Spectramax Emax (Molecular Devices) spectrometer for measuring absorbance with five filter choices wavelengths in the visible wavelength (400nm-750nm). These provide an opportunity to perform ELISA, BCA for total protein content and various other end point assays. There is a Gemini XPS spectrofluorometer, which allows with filters required for most fluorescence-based assays. Cellular and molecular binding studies, DNA quantitation, drug screening, and SNP genotyping analysis all require a high degree of sensitivity can be performed. Both plate readers are accessed through SoftMax® Pro Software, loaded on to a dedicated computer.
There is a YSI-2700 Biochemistry analyzer (YSI Life Sciences, Yellow Spring, OH), that could be used in various analyses including glucose.
Spectrum of electrophoresis equipments XCell SureLock™ Mini-Cell Electrophoresis apparatus (Invitrogen) for DNA/RNA analysis and western blot analysis modules. There is an HPLC system (Dionex Co., Sunnyvale, CA, USA) containing a Waters Symmetry 300 column (4.6 mm × 150 mm dimensions, 5 μm and 90 Å packing). There is a Dispersion Zeta Potential (Model DT-1200) meter for measuring particle sizing and charge density. There is a Q2000 differential scanning calorimeter (TA instruments New Castle, DE) equipped with refrigerated cooling system (RCS) 90. There is a Thermo-Nichole-50 FTIR spectrometer equipped with a DTGS detector.
There is a LCR Databridge (Instek, CA, USA) with RS-232C Interface (Includes LCR-Viewer Software) is also available in the laboratory through which allow 2 Hz to 200 kHz, Continuously Variable frequency current at 0.05% accuracy. Test condition and test result shown on the Screen Simultaneously. There is also a ES30P-5W/DAM Gamma High Voltage Research (Ormond Beach, FL) equipment useful for electrospinning.
Cell Culture
- Tissue Culture
Madihally is the primary custodian and operator for a tissue culture facility to culture mammalian as well bacterial cells. These facilities are equipped with two Thermo Forma Model 1284 Class II Type A3/B Biological Laminar Flow Hoods with sliding windows. There is one CO2 controlled water-jacketed Forma™ 3110 incubator for routine cell culture experiments. There is another Forma™ 3110 incubator O2 & CO2 controlled water-jacketed Incubator, which allows performing experiments at hypoxic conditions. In the same room, there is a 3-L temperature controlled centrifuge, which helps in cell harvesting routinely from the cell culture plates. In the same room, there is a refrigerator so that all cell culture supplies can be stored. There is a Locator Jr. cryo biological storage system to store cells and other components in liquid nitrogen.
- Flow Cytometer
Madihally is the primary custodian and operator for a Becton Dickinson FACSCalibur flow cytomter. It is not a part of the core facility. It is equipped with a low-power (15mW) argon laser that emits light at 488 nm as well as a low-power helium-neon diode that emits light at ~635 nm. It has two detectors for light scatter and three detectors for fluorescence. The optical filters on this cytometer are not interchangeable and the choices of fluorophores is constrained. These are accessed through a dedicated computer loaded with Cellquest software.
Recently, BD FACSAria III Cell Sorter was purchased through a grant from OSU Facility Renovation and Core Facility program. Madihally is a co-PI on the grant and this expands the capabilities of OSU’s Flow Cytometry, which also has couple of other flow cytometers. Madihally is a board-member of the facility. If necessary, cell sorting can be performed.
Microscopy
There are four different microscopes available in the laboratory which are listed below. Apart from these, there is also a multi-user microscopy laboratory which is available for all Faculty members and graduate students on a per hour cost basis. The equipment includes an atomic force microscope, a confocal microscope, a transmission electron microscope, Skyscan-1074 microComputer Tomography (in CHS), microtome, cryomicrotome, and embedding counsel
- Nikon TE2000-U Microscope
This is an inverted microscope fitted with high quality 4, 10, 20, and 40X objective lenses with phase contrast options. There is also Epi-fluorescence capabilities with different filters such as ultraviolet, blue, and green which enables collection of fluorescent images. High resolution black and white images can be captured with Hamamatsu CCD Camera with an internal cooler (C4742-95-12ERG). Low resolution color images can also be captured using digital color camera. There is also an Image Analysis Software “Sigma Scan Pro” apart from ImageJ, loaded on a dedicated computer with 1tB hard drive.
- EVOSTM AME-i2111 Digital Inverted Microscope
It is a fully integrated microscope with a digital camera and LCD display. Since it occupies very less floor space, it is kept in the tissue culture facility and allows easy view, and capture images quickly without the need of transporting cultures to another location. It has 4x, 10x Ph, 20x Ph Objectives. Stage has full range of motion in X and Y directions. Stage is manually controlled. Image data are stored on a computer connected to the microscope and accessed through Micron software, loaded on a netbook which occupies very little space. Digital images are then processed for quantification or publication in the computer connected to Nikon microscope.
- Olympus Provis AX70 Microscope
This microscope incorporates multiple illuminators (episcopic and diascopic), analyzers, DIC prisms, fluorescence, and phase contrast capabilities. It has 1.5, 2, 4 and 20X objectives. The micrography system is the ultimate in sophistication and performance featuring spot measurement, automatic exposure control, and zoom magnification for flexible, easy framing.
- Joel JSM-6360 Scanning Electron Microscope
It is operated under high vacuum. Up to 3,00,000x magnification is possible with a resolution of 3 nm. Accelerating voltage come go up to 30 kV. It is interfaced with a computer which is also used to control the function.
Miscellaneous Equipment
There are four variable speed Masterflex L/S peristaltic pumps (Cole-Parmer, Vernon Hills, IL), with two different pump heads, allowing the possibility of running bioreactors at various speeds. Further, these pumps are controlled through a Masterflex software loaded on to the computer connected to the pump. There are two flow-through oxygen electrodes (16-730, Microelectrodes, Inc., Bedford, NH), and one in-line physiological pressure transducer (Capto SP844, Skoppum, Norway). Data from oxygen electrodes and pressure transducer are acquired using Chart™ 5 for Windows via the Powerlab/4SP System (ADI Instruments, Inc., Colorado Springs, CO). These instruments are used in the bioreactor operations to monitor respective parameters.
There are two syringe pumps (74900 series, Cole-Parmer Instrument Company, Vernon Hills, IL) that can be used to pump fluids at required flow rate for use in electrospinning.
There is 6-L Vertis bench-top Freezemobile 6.6 Series System in the laboratory, equipped with a smooth walled corrosion resistant stainless steel condenser -85C with 140 in2 of cold surface area. (Capacity is 3.3 L per 24 h). This system is connected to 255 L/min vacuum pump with corrosion resistant components and adjustable gas ballast.
There are numerous analytical balances, rotary/linear Shaker, circulating reverse osmosis deionized water, large steam sterilizer, miscellaneous durable items (e.g. pH meter, water bath, magnetic stirrer, pipetters, vacuum desiccators etc.), multiple fume hoods, sinks, storage cabinets (independent acid, base, corrosive and flammable cabinets) are present in the laboratory.