Automated Epifluorescence Microscopy and Analysis Workstation; ImageXpress Micro by Molecular Devices
The ImageXpress Micro ™ cellular imaging system by Molecular Devices is a fully-integrated hardware and software system for high throughput automated epifluorescent cell-based imaging. The SCC Shared Facility’s system has environmental control of temperature and 5% CO2 and single-channel pipetting for kinetic experiments. The Shared Facility also has licensed the MetaXpress® and MetaMorph cellular image analysis software for high content analysis. Has 2x, 4x, 10x, 20x, 40x, and 60xoil objectives, and filter sets for DAPI, CFP, GFP, FITC, YFP, TRITC, Cy3, TxRed, Cy5, FRET, and a filter cube optimized for visualization of alkaline phosphatase product.
Photon Microscopy; Prairie Technologies
In multi-photon mode there are 2 PMTs for simultaneous acquisition from a single high powered ultra-fast (femto-second pulse) Ti:Sapphire Chameleon Ultra II laser (Coherent). Z-series, T-series, and targeted area laser exposures for FRAP, FRET, ablation, photoactivition, and photobleaching are easy to setup, as well as precise X-Y coordinate automated image acquisition. The Olympus BX51WIF is an upright microscope specially optimized for water immersion microscopy techniques. Selected objectives give high quality images for tissue slices, cell colonies, or monolayers of cells on different bioengineered materials or traditional culture dishes and slides. The advantages of two-photon excitation have allowed experiments to be performed that would not be possible using confocal microscopy.
Confocal Microscopy; SFC by Prairie Technologies
The Swept Field Confocal (SFC) is a Prairie invention that brings both high spatial resolution and high temporal resolution together into a single compact confocal scanner. Users can control the acquisition time and laser exposure with software control, as well as the frequency and duration of each “sweep” of the scanner. Highest spatial resolution is obtained by using the pinhole size best suited to a given objective lens. Z-and T-series are easily set up with scan speeds in excess of 30 frames per second (fps). Confocal laser lines currently in place are 405, 488, 561, and 643 nm.
Bio-Plex MAGPIX Multiplex Reader
The Bio-Plex MAGPIX multiplex reader is a compact, robust system for magnetic bead–based immunoassays. This multiplex reader is capable of reading assays designed on magnetic xMAP (MagPlex) beads. Read up to 50 analytes per sample (as little as 25 ul), and can be used to quantify levels of phosphoproteins and cytokines that are present in cell or tissue lysates, serum or plasma samples. It can also be used to analyze both proteins and nucleic acids. Researchers can quickly evaluate analytes by using pre-validated assays available in the areas of cancer, cardiovascular, cell signaling, cellular metabolism, immunology, metabolism and endocrinology, neuroscience, stem cells, and toxicity. Researchers can quickly evaluate analytes by using pre-validated assays available in the areas of cancer, cardiovascular, cell signaling, cellular metabolism, immunology, metabolism and endocrinology, neuroscience, stem cells, and toxicity.
Perfusion and Heated Stage Platform for 2P and Confocal Microscopy by Warner Instruments/Harvard Apparatus Company
5% CO2 can be delivered to the medium that will be perfused across the live sample that is either in an open configuration (QE-1) for direct access to the Prairie Technologies 2-Photon/SFC upright microscope system, or enclosed between glass coverslips (Quick Change). The Six Channel Perfusion Valve Control Systems (VC-6) setup is for medium to low flow, and allows easy change over between different media compositions during imaging.
Flow Cytometry; FC 500; Beckman-Coulter
The Beckman-Coulter FC-500 is a user-friendly analytical instrument with two laser excitation wavelengths produced by a 488nm argon laser and solid-state 25mW red diode laser operating at 635 nm. Researchers can measure up to 5 colors simultaneously.
This cytometer operates as a ‘walk-away’ system. It has a carousel that may be loaded with up to 32 tubes each to be run automatically. This instrument is used for multiparameter immunofluorescence detection, stem cell type verification, cell cycle, apoptosis, green fluorescence protein expression.
Parameter Spectra Fluorochromes
|488nm||FL1||525nm||FITC, AF488, GFP, YFP, CFSE, Rhodamine123, Vybrant Green|
|488nm||FL2||575nm||PE, DsRed, Vybrant orange|
|488nm||FL3||620nm||PE-Texas Red(ECD), PI|
|FL4||675nm|| PE-Cy5, PerCP, 7AAD
APC, AF 633, AF647
|635nm||FL5||755nm||PE-Cy7, APC-Cy7, APC-AF750|
FlowCytometry; Guava EMD/Millipore
The Guava EasyCyte 6HT is a benchtop flow cytometer with 2 excitation sources at 488 nm and 532 nm, one forward scatter diode detector, and 5 emission PMT detectors: side scatter SSC, 510/20, 583/26, 610/20, and 690/50. Also, phycoerythrin type markers will be more highly excited and more easily detected with this laser rather than the 488 nm laser alone. The Guava offers a built-in 96-well autosampler, which allows researchers to obtain even more data from small sample sizes than current systems, while performing complex biological studies such as cell signaling, human and mouse embryonic stem cell differentiation, and multiplex compound screening. A strong attribute of this instrument is that a commonly used fluorescent protein, mCherry, is detectable using this instrument due to the inclusion of the 532 nm laser. Monitoring the interplay of up to six different biological signals simultaneously improves the efficiency of experimental outputs. The Guava easyCyte 6HT-2L System runs all Guava Assays and uses Guava InCyte software for data acquisition and analysis. Guava InCyte offers drag and drop gating, pre and post acquisition compensation, heat map display, and IC50/EC50 curve generation.
Stem Cell Culture Room
Equipped with 3 BL-2 laminar flow Hoods, 4 CO2 incubators, centrifuges, epifluorescent/phase/color Zeiss inverted microscope, Evos xl core microscope, water bath, -20C freezer, hemacytometer and Scepter, GentleMACs cell dissociator, and storage space (each lab keeps their own stock of supplies).
Chemical Synthesis Hood With Photo Resist Spinner
Vertical Laminar Flow Clean Air Wet Process Fume Hood; Spinner has a microprocessor sequence controller for rpm, ramp, time, and accessory control for dispense and exhaust.
Electroforce 3200 by Bose Electronics
The ElectroForce® 3200 test instruments was optimized for the characterization of Native Tissues, Scaffolds, and Biomaterials. It features a 225 N maximum force. With the versatility of static to 200 Hz frequency response, the table-top configuration is adaptable to a variety of biomedical research and engineered materials test applications, including torsion testing, creep under dynamic loading and special environments (hot/cold chambers). We have one set of test grippers optimized for our 5 lb load cell, and we also have a 250 g load cell for which accessories may be ordered or expertly machined.
The SSCF SEC-MALS system combines Size-Exclusion Chromatography (Agilent LC 1260) with Multi-Angle Light Scattering (Wyatt Dawn Heleos II) and Refractive Index Detection (Wyatt Optilab T-rEX). The Agilent HPLC instrument is capable of separating polymer and protein mixtures on the basis of size and can provide compositional information on the basis of UV absorbance measurement. The in-line light scattering instrument can quantify the size of the separated polymers or protein measurements with molecular weights and radii of gyration. Finally, the refractive index detection measurements can be combined with the other data collected by the system to simultaneously measure the molecular weight distribution of a polymer and the extent to which it has been conjugated by functional proteins/peptides. This technique was developed as a result of a Healy/Schaffer collaboration, and it is essential to evaluate the valency of multivalent conjugates. For instance, students in the Healy and Schaffer labs have been using these conjugates to optimize the differentiation protocols of ES and iPS cells and to improve the efficiency of these procedures. The SSCF allows interested researchers to use this equipment individually or in combination to characterize their polymer/peptide conjugates.