Equipment
Please refer to iLab for pricing
Benchtop Analyzers
- 3 lasers – blue (488nm – 50mW), violet (405nm – 100mW) and red (640nm – 80mW)
- 2 scattering channels
- up to 38 fluorescence detectors
Additional Resources
Optimized Multicolor Immunofluorescence Panel (OMIP) is a special peer-reviewed Cytometry Part A publication type that reports on newly designed and optimized multicolor panels for flow cytometry.
Click here for a panel design grid to help you design antibody panels.
Click here to evaluate and optimize your panel design using the Cytek Full Spectrum Viewer.
- 5 lasers – blue (488nm – 50mW), violet (405nm – 100mW), yellow-green (561nm – 50mW), red (640nm – 80mW) and UV (355nm – 20mW)
- 3 scattering channels
- up to 64 fluorescence detectors
- Autosampler – 96 well plate
Additional Resources
Optimized Multicolor Immunofluorescence Panel (OMIP) is a special peer-reviewed Cytometry Part A publication type that reports on newly designed and optimized multicolor panels for flow cytometry.
Click here for a panel design grid to help design your antibody panels.
Click here to evaluate and optimize your panel design using the Cytek Full Spectrum Viewer.
- 4 laser lines – blue (488nm), red (640nm), violet (405nm), and yellow-green (561nm)
- 16 parameters (FSC, SSC and 14 fluorescent parameters)
- Incorporated high-resolution, high-speed brightfield microscope capable of capturing up to 6,000 images/second
- Autosampler – 96-well, 384-well, and deep-well plates
Click here for Attune CytPix Optical Configuration.
- 4 laser lines – blue (488nm), red (640nm), violet (405nm), and yellow-green (561nm)
- 16 parameters (FSC, SSC and 14 fluorescent parameters)
- Autosampler – 96-well, 384-well, and deep-well plates; 1.5 mL and 2 mL microcentrifuge tube rack (up to 24 per rack); Passive cooling available for 96-well U-bottom plates and microcentrifuge tube racks
Click here for ATTUNE NxT Optical Configuration.
- 4 laser lines (488nm, 405nm, 561nm, and 640nm)
- 18 parameters; 2 scatter, 3 colors off the blue laser, 5 colors off the yellow-green laser, 3 colors off the red and 6 off the violet.
Click here to access the optical layout.
Plan your panel for the LSRFortessa here.
All individuals who desire training must first complete the DIVA 6.0 Online Tutorial (even those with previous DIVA experience) as the training will assume a basic knowledge of the software.
See Scheduling for information on scheduling time on this instrument.
- 3 lasers – blue (488nm) violet (407nm) and red (637nm)
- A total of five channels can be selected; 3 off of the 488nm and either 2 off the 407nm or 1 off the 407nm and 1 off the 637nm.
Click here to access the optical layout.
Panel design assistance can be found here (Blue/Violet) (Blue/Red/Violet).
The Calibur uses CellQuest Pro and Rainbow Acquisition software. Please review the Rainbow Video Tutorial before using.
A flexible analyzer based on the principles of flow cytometry enables you to multiplex (simultaneously measure) up to 50 analytes in a single microplate well, using very small sample volumes. The system delivers fast and cost-effective bioassay results on many assay formats including nucleic acid assays, receptor-ligand assays, immunoassays and enzymatic assays.
-
- For a list of companies who supply reagent kits for use on the Luminex platform click here.
- For a list of publications for this technology click here.
- For a list of applications and updates for this technology click here.
- For a link to general protocols try this xMAP Luminex Cook Book
This instrument is located in Pinn Hall, Rm 2011. 434-243-2711
Cell Sorters
Like the Aurora full spectral cytometers, the Aurora CS provides the benefits of full spectral cytometry. Easily transition of already optimized high-dimensional fluorescent panels directly from existing Aurora Spectral Cytometers to the Aurora CS Cell Sorter. Ability to perform high purity cell sorting on complex cell subsets and recover high-dimensional analytical data above 40+ fluorescent markers, maximizing data generated on valuable limited samples.
- Capable of detecting up to 64 fluorescent parameters and 3 scatter parameters
- 6-way sorting 1.5ml tubes
- 4-way into 5 ml tubes
- 96 well plate sorting
- BSC Class A2 level II biosafety cabinet for sorting BSL-2 biohazardous samples
- 5 laser lines – UV (355nm), violet (405nm), blue (488nm), yellow-green (561nm), and red (640nm)
- 17 parameters – forward & side scatter and 15 fluorescence detectors
- Six-way sorting into tubes or single cell sorting directly into plates or slides
- BSC Class II biosafety cabinet for sorting BSL-2 biohazardous samples
Consult the facility for available filter set ups, fluorochrome combinations, and appropriate controls, prior to designing your experiment.
For a current optical layout and possible fluorochromes please click here.
- 4 laser lines – violet (405nm), blue (488nm), yellow-green (561nm) and red (638nm)
- 14 parameters – forward & side scatter and 12 fluorescent detectors
- Two and four-way sorting into tubes as well as 6-, 12-, 24-, 48-, 96-, and 384-well plates.
- BSC Class A2 level II biosafety cabinet for sorting BSL-2 biohazardous samples.
- Automated setup
- Isolation of cell populations labelled with 50nm superparamagnetic particles
- Choice of manually labeling cells offline or having the autoMACS Pro label them for you as part of the isolation protocol.
- Several different automated protocols based on:
- Density of the antigen
- Frequency of the population
- Negative or positive selection
- Isolation directly from whole blood
AutoMACS Pro Time Per Sample Calculator
Mass Cytometer
The CyTOF XT Mass Cytometer simultaneously resolves multiple parameters with minimal signal overlap, providing unprecedented insights into the functional and phenotypic complexity of biological systems at the single cell level.
The CyTOF XT analyzes cells labeled with a panel of reagents conjugated to heavy metal isotopes using state-of-the art time of flight mass cytometry technology.
With over 100 detection channels and over 30 isotopic metal probes, CyTOF XT simplifies panel design and unlocks the functional and phenotypic complexity of biological systems at the single cell level.
Mass cytometry experiments routinely measure dozens of functional and phenotypic parameters in millions of single cells, output as .txt and .fcs files. Analysis of such high dimensional datasets is performed using a tool kit that includes not only traditional histograms and bivariate plots, but also novel clustering algorithms, neural networks, and dimensionality reduction methods.
CyTOF data analysis – High-Dimensional Single Cell Analysis
Consult the facility for metal combinations, appropriate controls, and other considerations prior to designing your experiment.
Located in Pinn Hall, Room 2013; 434-243-7682.
Imaging Flow Cytometer
By combining the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insight of microscopy, the ImageStreamX overcomes the limitations of both techniques and opens the door to an extensive range of applications: follow translocation from the cytoplasm to the nucleus; quantify capping and co-capping reactions; separate cells based on unique morphologic criteria; study cell-cell interactions; differentiate cell states such as late apoptosis and necrosis, study cell signaling, co-localization, shape change, internalization… – the range of potential applications is extremely broad.
You can define and quantify cell subpopulations not only by signal intensity, but by the location of fluorescent probes on, in, or between cells, even in rare sub-populations and highly heterogeneous samples. You can use cell and nuclear morphologies to characterize cell populations.
In the ImageStreamX, cells are illuminated with both a brightfield source and 405nm, 488nm, 560nm and 658nm lasers. It produces up to 12 high-resolution images (brightfield, darkfield and up to 10 fluorescence parameters) of each cell directly in flow, at rates up to 5,000 cells per second.
The IDEAS software package’s numerous pre-defined fluorescence and morphologic parameters let you quantitate virtually anything you can see so you can take full advantage of complex visual data to perform statistically-robust, multi-parameter quantitative analyses.
Applications that require the enumeration of spots within the cell or nucleus, like fluorescence in situ hybridization (FISH), require an increased depth of field. The ImageStreamX’s Extended Depth of Field (EDF) upgrade is a combination of specialized optics and unique image processing algorithms that project all structures and probes within the cell into one crisply-focused image. EDF makes it possible to find and count spots anywhere within the cell without any sacrifice in sensitivity or speed.
New features:
- The Mark II upgrade provides real-time compensation, plotting, gating, and the use of gated events as data storage criteria. It accepts sample volumes from 20 to 200μL with 95% sample utilization thanks to its new fluidic system. It also sports a redesigned, intuitive user interface.
- The new MultiMag option provides 20X and 60X objective lenses in addition to the standard 40X lens for greater experimental flexibility and improved resolution. The 60X objective increases magnification for small objects such as yeast and bacteria and offers greater detail with mammalian and plant cells. The 20X objective provides an increased field of view for the imaging of large cells, such as epithelial cells.
- The 96-well plate AutoSampler option for the ImageStreamX imaging flow cytometer enhances productivity with unattended sample loading from multiwell plates. The fully integrated AutoSampler option allows you to easily perform dose response and time course studies even in primary samples, thanks to the ImageStreamX’s ability to image large numbers of cells from each sample.
Click here for a full equipment list across all School of Medicine Research Cores