The FGCF houses of a variety of equipment to support research needs on campus, ranging from PCR amplification to microscopic imaging. To get information about each instrument select the appropriate drop down menu. For questions about specific equipment or applications, please contact FGCF coordinator Ryan Hanson.
PCR systems
96-well format thermal cycler with standard temperature range from 4-99掳C. Sign-up sheet placed next to instrument.
Potential applications:
- PCR amplification of nucleic acid templates for plasmid design, genotyping or sequencing.
ABI Veriti thermal cycler with 96-well format. Simple touch screen control for setting up cycling programs. Capable of heating gradients for optimizing PCR conditions.
Potential applications:
- PCR amplification of nucleic acid templates for plasmid design, genotyping or sequencing.
QuantStudio6 Flex Real-Time PCR instrument with both 96 and 384-well formats available. Multiplex capabilities including FAM, NED, ROX, SYBR and VIC. Touch screen and software based set-up and data export.
Potential Applications:
- Analysis of gene expression through both single and multiplex assays
- Absolute quantifications with standard curves
- Melt curve analysis of primer design
- Genotyping.
Spectrophotometers
UV/Vis spectrophotometer for the quantification of DNA, RNA and protein samples using small volumes (0.5-2.0 ul). Detection range of 2 ng/ul-15,000 ng/ul for dsDNA and 0.1 mg/ml-400 mg/ml for BSA. Xenon flash light source with wavelengths spanning 190 nm-840 nm.
Potential applications:
- Quantification of DNA from plasmid preparations, genomic isolation and PCR reactions.
- Measurements of RNA concentration.
- Quantification of protein concentrations.
BioTek Synergy2 microplate reader is a multi-modal UV/Vis spectrophotometer capable of absorbance, fluorescence and luminescence based assays for standard multi-well plate formats (6 to 384-wells). FGCF system has the dual reagent dispenser module for controlled addition to compounds during analysis. Contact FGCF manager for filter sets available for use.
Potential applications:
- Protein quantification via Bradford.
- Cell viability assays via absorbance, fluorescence and luminescence.
- readouts of transcriptional reporters through fluorescence and luminescence.
Gel Imaging
Licor Odyssey Fc imaging system equipped with 600, 700 and 800 nm scan channels. Capable of dual color imaging of western blots in the 700 and 800 channels. 600 nm channel for detection of DNA stained with the SYBR family dyes. Image Studio acquisition software can be used to quantify band intensities for comparisons of individual lanes.
Applications:
- Western blot imaging
- DNA gel imaging
- Analysis of gel images
Microscopes
Olympus stereomicroscope for low-level magnification of tissues, organisms and materials. Magnification wheel from 0.7-11.5X along with a 10X magnification eyepiece. This microscope is set up for both transmitted and reflected light microscopy. Additionally, we have an LED-light source for epifluorescence imaging using FITC and RFP filter cubes. Camera is interfaced with a DP71 digital camera and cellSens for image acquisition.
Potential Applications
- Dissection or manipulation of fine tissues.
- Fluorescence imaging of tissue or transgenic organisms.
- Imaging of surface details by reflected light microscopy.
The Olympus Bx53 upright microscope is equipped with 5X, 10X, 20X and 40X air objectives, as well as a 100X oil-immersion objective for high magnification imaging. The Bx53 microscope is equipped for DIC imaging with the 10, 20 and 40X objectives and phase contrast with the 100X objective in addition to standard Brightfield capabilities. This microscope is also equipped with the X-Cite LED illumination source for epifluorescence with DAPI, GFP, YFP and TRITC filter cubes. For image acquisition this microscope is interfaced with a DP80 digital camera and cellSens image acquisition software.
Potential Applications
- Brightfield imaging of histological slides.
- Phase and DIC imaging.
- Fluorescence imaging of DAPI, GFP and RFP based-fluorophores.
The Olympus IX70 inverted microscope is readily equipped for Brightfield, phase contrast and epifluorescence imaging. This microscope is equipped with 10, 20 and 40X air objectives. The inverted objectives allow for easy access to samples making it ideal for imaging cells in plates or dishes. A mercury lamp source is used for epifluorescence imaging coupled with DAPI, GFP and RFP filters. A DP73 digital camera captures images in conjunction with the cellSens acquisition software.
Potential Applications
- Brightfield and Phase imaging of cells in plates or dishes.
- Fluorescence imaging of cells in culture to validate treatments or transfections.
The Olympus FV1200 system provides high-resolution imaging and optical sections through pinhole rejection of out of focus light. The FV1200 system is equipped with 405/458/488/515/559/635 laser lines to excite the majority of available fluorophores and fluorescent proteins. For emissions detection, the system is equipped with 2 variable band-pass filters to enable flexible emission collection for the first two detectors. The system also has a third detector with a standard bandpass filter for up to 3-channel simultaneous imaging. A transmitted light detector also enables the collection of Brightfield images. 4X, 10X, 20X and 40X air objectives, as well as a 60X oil-immersion objective offer a range of magnifications for imaging. Inserts for both slides, dishes and multi-well plates.
Potential applications
- Optical sectioning and three-dimensional imaging.
- Spectral unmixing and autofluorescence identification with lambda scan.
- short-term time lapse imaging.
The Leica LIAchroic Stellaris 5 confocal system provides high-resolution imaging through pin-hole rejection of out of focus light and can further increase resolution using the included Lightning deconvolution software, resulting in super-resolution imaging of samples. This system is equipped with 405/488/514/559/638 laser lines for excitation of most fluorophores and fluorescent proteins. Available objectives include 5X, 10X, 20X air objectives and both 40X and 60X oil-immersion objectives for high-resolution imaging. The scan head is equipped with 3 HyD spectral detectors for flexible collection of emissions. The system is also equipped with a stage-top incubator for environmental control enabling longer-term live cell imaging. Improved image acquisition speed makes the Leica ideal for imaging large Z-stacks or performing large imaging stitching of multiple fields.
Potential applications
- High-resolution imaging and super-resolution with Lightning deconvolution.
- Long-term live cell imaging.
- Large image stitching.
Nikon Ti2 widefield fluorescence microscope equipped with an 8-line LED excitation system 390/440/475/510/555/575/635/747 to hit virtually all fluorophores and fluorescent proteins. Equipped with 10X, 20X, and 40X air objectives with a 0.95 NA on the 40X objective. Full-enclosure incubator for environmental control and live-cell imaging without interference from room lighting. Perfect focus system to ensure no focal drift over time. Polygon DMD for precise photostimulation or optogenetic approaches. Orca Fusion-BT sCMOS camera for image capture with large dynamic range and low noise. NIS-elements software equipped with JOBS for complex imaging experiments.
| Long-term imaging structure | |
|---|---|
Imaging duration | Rate |
0-4 hours | $15/hr |
4-24 hours | $60 daily maximum |
24-48 hours | $45 daily maximum |
48-72 hours | $30 daily maximum |
96+ hours | $15 per additional day |
Potential applications
- Live cell imaging for long durations.
- Imaging fluorescent proteins not ideal for confocal applications due to off-peak excitation.
- Photostimulation and optogenetics.
Histological prep
The Olympus CUT 4060E microtome allows users to cut and prepare their own slides from FFPE sections. Auto-sectioning feature allows for easy facing of block prior to sectioning.
Potential Applications
- Sectioning of FFPE samples
Leica CM1850 UV cryostat is equipped with a rapid-cool shelf for preparations of samples. Manual and automatic defrost programs can be used to minimize frost buildup. Electronic coarse feed is used to orient sample prior to cutting. Dial selector for easy to read section thickness and anti-roll plate to minimize curling of sections as they are cut. Cryostat is equipped with a CRYOVAC system for easy disposal of unwanted sections.
Potential Applications
- Cryosectioning for slides.
- Preparation of samples for spatial transcriptomics.
GC-MS and HPLC systems
Agilent 5975 Gas Chromatograph/Mass Spectrometer
Agilent High-Performance Liquid Chromatography system
Centrifuges
The Beckman OPTIMA 130K Ultracentrifuge is equipped with MLS-50, TLA-120.2 and MLA-55 rotors for high-speed centrifugation needs.
Additional Equipment
Additional equipment available upon on request includes the following. Users are expected to provide consumables.
- Agilent Bioanalyzer
- BioRad Gene Pulser Xcell Electroporation System
- BioRad PROTEAN IEF System with PROTEAN Plus Multi-Casting Chamber, PROTEAN Plus Dodeca Cell, PowerPac HC Power Supply
- Qubit 3 Fluorometer