Lab Facilities

Laboratory

Dinu’s laboratories encompasses 1050 square feet and are located in the Statler College's Advanced Engineering Research Building. The labs are in close proximity and fully operational. Some of the specific equipment is included below:

Electric Cell Impedance Sensing

The ECIS Zθ (Z theta) is an advanced, automated, non-invasive means to monitor cell behavior in real-time and without the use of labels. The ECIS Zθ is capable of automatically monitoring from 8, 16 or 96 tissue culture wells and returning quantitative information regarding the impedance of small 250 micrometer electrodes used as substrates for cell growth. The system can be run at different AC frequencies from 50 Hz to 100,000 Hz and monitors both the voltage and phase of AC signal across the active electrodes with data sampling as often as 25 points per second. 

Software included with the system interprets this information as both a resistance and capacitance (treating the active electrode with cells as a simple RC circuit). In addition to performing continuous time course acquisition of data, the instrument can also automatically scan 11 different AC frequencies to present data in the form of an impedance frequency spectrum. 

Included with the system are modeling programs designed to calculate cell parameters from the raw voltage and phase data. Cell parameters that can be followed in time include the barrier function (permeability) of confluent cell layers, membrane capacitance, and a term (alpha) related to the spacing between the cell’s basal membrane and the active electrode. 

The instrument comes with a dedicated personal computer with installed software that has been factory tested with the system and all required cables. ECIS Software is compatible with Windows XP, Vista and Mac OSX. Applied BioPhysics, Inc. is the only source of this type of instrumentation. 

Instruments that measure TER (transepithelial resistance) are designed for extremely tight epithelial layers and only make single point measurements; whereas, ECIS is not restricted to tight layers and can monitor any adherent cell type in monolayer or sparse culture. The ECIS Z-Theta system has an optional high field module that allows one to carry out automatic wound healing assays. 

There is no other source of this technology on the market today. There are no suppliers for this type of instrumentation. Instruments that measure TER (transepithelial resistance) are designed for extremely tight epithelial layers and only make single point measurements; whereas, ECIS is not restricted to tight layers and can monitor any adherent cell type in monolayer or sparse culture. 

The acquisition of this instrument is essential to the success of this award. As stipulated in the attached SEED proposal that was recently awarded, all the experiments to be performed explore the ECIS unique features. If this sole source justification is not approved no success can be assured for this award which in turn will lead to interruption of the funding. Such outcome will also lead to interruption of the funding for the student employed by this project.

MFP-3D-BIO

• MFP-3D ARC2 controller (operating modes: Contact, AC with Q-Control, Dual AC, Force Mode, LFM, MFM, KFM and MicroAngelo Nanolithography and nanomanipulation)
• MFP-3D high clearance scanner/stage, appropriate base plate for specified IO make and model, standard MFP-3D cantilever holder compatible with both air and liquids
• Computer specs: Dell Precision T3500 w/Windows XP Professional, Intel Quad Core Xeon 3500 series, (2.66GHz), 3GB DDR3 RAM, Two 320GB SATA Hard Drives (RAID1 mirror), 16x DVD+/-RW (fast dual-layer DVD writer), nVidia GTS 250 512MB video card, eight USB 2.0 ports, two2 FireWire ports, 525W power supply, two 20-inch LCD flat panel displays
• IgorPro 6 with Asylum Research data acquisition and control software, argyle analysis, image processing and rendering software
• Top-View IO feature provides viewing of the tip and sample from above through a 10X Mitutoyo objective, with brightfield epi-illumination, on inverted optical systems
• Low-coherence 860nm super luminescent diode source for optical lever deflection measurement. Low coherence enables flat-baseline, low-noise force and topography measurements. Incorporates a narrowband source filter to suppress out-of-band light emissions, eliminating crosstalk into fluorescence and other optical measurements.
• Phase Contrast Illumination: (Ph1, Ph2 and PhL/C/0) simultaneously with AFM by illuminating through the AFM head. Includes a custom optical subsystem, annulus slider and field diaphragm.
• Infrared Blocking Filter: eliminates infrared light from the MFP-3D super luminescent diode that would otherwise appear in sensitive optical measurements analog monochrome CCD camera kit, 1/2": for side port of the inverted optical microscope. Video Image is captured and displayed with MFP-3D software.
• Petri dish holder that supports the bottom of the Petri dish to limit “drumhead” vibrations and reduce noise.
• Sample package of assorted Olympus cantilevers/probes.
• Miscellaneous accessories: tweezers, calibration grating, static master, IR card, bubble level, earphones, headstand, power strip and all necessary cords and cables for operation of the AFM.
• BioHeater, temperature-controlled version: the BioHeater includes a resistive liquid immersion heating element and thermistor. Full top and bottom optical access. Software selectable temperature control of the cell can be maintained from ambient temperatures up to 80°C with a precision of 0.1°C and accuracy to 0.2°C.
• Petri dish heater: this modified version of the MFP-3D scanner top plate is designed to hold a variety of cell culture dishes and can be heated from ambient temperature to 45°C. Temperature can be controlled to within 0.1°C. Supported dishes are Willco Wells and WPI glass bottom and two polystyrene dishes by Falcon and Corning. The dish holder comes with a kit that includes means for magnetically securing the dishes to the AFM and an elastomer membrane to prevent evaporation. The dish holder is compatible with high NA 100X objectives. In addition to dishes, the dish holder still allows the use of standard microscope slides.
• Environmental controller: stand-alone temperature controller with bi-directional current supply for use with Asylum Research heating and cooling sample stages. Built-in microcontroller regulates closed loop temperature feedback independent from the AFM controller or PC computer. Fully operated through the graphical interface of the MFP-3D AFM software. Temperature controller along with resistive heating elements and thermistors provide temperature control. For use with Asylum Rresearch temperature-controlled stages.
• ORCA: Conductive AFM cantilever holder and accessory kits. Provides current imaging measurements at a constant applied voltage and point spectroscopy plots at user selected points. Kit includes: ORCA cantilever holder with integral pre-amp; standard gain is 5 x 10e8 volts/amp (~1pA to 10nA); conductive sample holder mount; 10-AC240TM Olympus electri-levers.
• iDriveTM kit for imaging in fluid: allows for magnetic actuation of special nitride probes exclusively offered by Asylum Research (Model BL-TR400PB). Kit includes iDrive cantilever holder and accessories. The holder can be easily configured in software to operate like a standard holder with piezo actuation for use with any “non-iDrive” probe.
• AVI-350 Vibration Isolation System with 24” x 24” x 3” granite top (BIO Systems); ideal isolation system for MFP-3D-BIO Systems; active 1.2 Hz to 200 Hz, passive > 200 HZ.
• Air Temperature Control (for BCH-45 Enclosure): forced air temperature control system installs inside the BCH -45 acoustic enclosure. Controls temperature by slightly heating (typically 3°C) the air surrounding the AFM. Typically the system will isolate the AFM from room temperature by 25X, leading to a comparable improvement in thermal drift.
• TMC Acoustic Noise Enclosure with Window designed for the MFP-3D-BIO or MFP-3D-CF: provides acoustically quiet environment for low-noise AFM imaging in a typical lab environment.

High resolution camera/ Microscope

• Photometrics System, Cascade 2:E2v CCD97: 16 Bit, 10 MHz digital monochrome camera with charge multiplication gain capability; liquid cooled with COOLCUBE.

• Nikon NIS Elements software: acquisition and analysis package, includes camera and Nikon motorized microscope drivers. Options modules include, advanced interpreter, live compare, time measurements, image database and 4D acquisition.
• NIS-Elements module: 4D imaging acquisition module for up to 4 dimensions, including any two of the following, time, channel or wavelength and focus.
• NIS-Elements module: time measurement.
• Nikon Ti-U microscope configured for Asylum AFM, TI-U inverted microscope main body, objectives (10x, 40x, 60x, 100x, NA 1.4), epi-fluorescence.

Core facilities available in the Statler College

The Statler College houses several shared facilities dedicated to nanotechnology research. These facilities include field-emission electron microscope and TEM with energy dispersive spectroscopy , inductively coupled plasma-optical emission spectroscopy, high resolution X-ray photoelectron spectrometers, atomic force microscopy, cytoviva hyperspectral microscopy, X-ray diffraction spectrometers, solid state NMR, mass spectrometers,and several Raman and FTIR spectrometers for surface chemistry characterization.

Core Facilities Available at WVU Health Sciences Center

1. Microscope Imaging Facility
   

   The facility provides access to advanced biomedical imaging technology including state-of-the-art microscopes and image analysis software packages. Equipment in this facility include Nikon Live Cell TE2000S microscope, Nikon Swept field microscope, Olympus Histology AZ70 microscope, Zeiss LSM 510 confocal microscope, Zeiss PALM MicroBeam system and Zeiss Violet LSM 510 confocal microscope.

2. Flow Cytometry Core Facility
   

   The facility is equipped with Becton-Dickenson FACSCalibur and FACSAria along with a Miltenyi Biotec AutoMACS magnetic bead cell sorting system. The Core has several software packages including CellQuest Pro, ModfitLT and BD Diva software for data acquisition and analysis.

3. Genomics Core Facility
   

   The facility performs and runs sequencing reactions and fragment analysis on an ABI model 3130XL genetic analyzer. The core also has an AlphaImager UV/Vis Image Documentation system, an iQ5 Real-Time PCR system, a Li-Cor 4300L DNA Analyzer and a ND-1000 NanoDrop spectrophotometer.

4. Pathology and Histology Facility
   

   The facility offers various services including autopsy service, surgical and cytopathology analysis. A centralized histology laboratory also provides routine processing, sectioning and staining of cell/tissue samples, as well as a variety of advanced immunohistochemical procedures.

5. Bioinformatics and Biostatistics Core Facility
   

   The core provides services on microarray design and analysis, 2-D gel electrophoresis, mass spectrometry data analysis, networks analysis, biostatistics, programming support, high-performance computing, sequence analysis and comparison, molecular dynamics and database design and implementation.

6. Animal Models and Imaging Facility
   

   The Animal Models and Imaging Facility has been established to accommodate the growing animal management needs of research laboratories and to provide state-of-the-art molecular imaging to WVU researchers and their collaborators. Conveniently located within the OLAR Animal Facility in WVU HSC, the AMIF currently performs acute and longitudinal bioluminescence and fluorescence studies on mice using the Xenogen IVIS® Lumina Imaging System. The system allows researchers to perform in vivo imaging using real-time imaging to monitor and record cellular and genetic activity within a mouse or rat using fluorescent and/or bioluminescent reporters. In addition, this facility also offers additional services such as mouse colony management, tissue collection, cryopreservation of sperm and various surgeries. The AMIF is directed by Karen Martin.