Flow Cytometry
Flow cytometry allows you measure many features of individual cells as they pass single-file through a laser beam, and physically isolate the exact populations you want to study.
The Flow Cytometry Shared Resource gives researchers access to a full range of cell analyzers and high-speed cell sorters, along with staff expertise to help design panels, run experiments, and interpret the results.
New users need to complete the Introduction to Flow Cytometry class, offered free by the facility. The class is held the first week of each month for three hours and covers the principles of flow cytometry technology, its applications, and how to use the facility.
New Project Consultation
Starting a new project involving one or more Shared Resources, or interested in learning more about available services? Contact Dr. Kate Hyde to schedule a consultation or complete the consultation form in Stratocore PPMS.
Request Service
Use Stratocore PPMS to request core facility services and manage scheduling and billing.
Pricing Benefits
Subsidized pricing is available to Fred & Pamela Buffett Cancer Center members. If you haven't received an email about subsidized pricing, please get in touch with cancer center administration to see if you qualify.Contact: buffettcancercenter@unmc.edu
If you conduct cancer-related research at UNMC and are not yet a member of a Cancer Center research program, visit the Fred & Pamela Buffett Cancer Center Membership page to learn more about becoming a member.
Ways to Work with the Flow Cytometry Shared Resource
| Option | What It Means |
|---|---|
| Full Service | Facility staff run your samples and return the data to you. |
| Supervised Use | You operate the instruments during core hours (Monday–Friday, 9 a.m.–6 p.m.), with staff available to assist as needed. |
| Independent Use | Once trained, you have 24/7 access to the analyzers. |
| Cell Sorting | Always performed by facility staff. Sorting is not available for independent use. |
| Consultation | Free at any stage—panel design, controls, troubleshooting, or interpreting your results. |
Instruments
Cell Analyzers:
Analyzers measure the cells in your sample but do not recover them. Spectral analyzers capture the full color spectrum of every dye rather than reading a handful of fixed color channels, then separate the overlapping signals mathematically. This makes it possible to run larger panels and to use dyes with similar colors that a conventional analyzer could not tell apart.
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Invitrogen Attune Xenith: Equipped with six lasers and 51 detectors, this analyzer allows both spectral and conventional analysis and is well-suited for high-parameter cell analysis.
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BD LSRFortessa: Five lasers detect up to 28 markers at once, well suited to deep immune profiling. Plate loader for 96- and 384-well screening.
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Thermo Fisher Attune NxT: Four lasers, up to 16 markers, with an autosampler for 96-well, 384-well, and deep-well plates. A strong choice for high-throughput work.
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BD LSRII: Four lasers, up to 14 markers, equipped with a plate loader for automated high-throughput of 96- and 384-well plates.
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BD LSRII Y/G: Six lasers, up to 18 markers, with a plate loader. A flexible option for demanding multi-color panels.
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Cytek Northern Lights: A compact spectral analyzer with one laser, detecting up to nine markers at once.
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A BD FACSCalibur: Two-laser (488-nanometer and 635-nanometer) flow cytometerable to analyze up to four-fluorescent parameters, as well as forward and side scatter.
Cell Sorters:
Sorters do everything an analyzer does, then physically separate the cells you have identified into tubes or plates so you can grow, sequence, or otherwise study them. All facility sorters are enclosed for safe handling of biohazardous (BSL-2) samples.
- Thermo Fisher Bigfoot: Five lasers and 53 detectors give this instrument the facility’s largest sorting capacity. Collects up to six populations at once into tubes from 1.5 to 50 mL, or plates up to 1536 wells, and runs using either conventional or spectral analysis.
- BD FACSAria: Five lasers, up to 14 markers. Collects four populations at a time, down to a single cell per well, into tubes or culture plates.
- Miltenyi autoMACS Pro: Separates cells tagged with magnetic beads by passing them through a magnetic field. Faster and gentler on cells than a sorter, bead separation is designed to enrich a cell type in bulk rather than isolate a precisely defined population.
Coming Soon! Spectral Imaging Flow Cytometry:
- FACSDiscover instruments are being added to the facility, both combining full-spectrum analysis with imaging of every cell—capturing where signals sit inside each cell, not just how much is present.
Analysis Software:
- FlowJo, BD FACSDiva, ModFit LT, and CellQuest Pro are available at no charge on facility workstations.
- UNMC researchers can also join the facility’s FlowJo site license (approximately $300 per year) to analyze data from any computer.
- Assisted data analysis is available if your dataset is complex or you are new to flow cytometry.
Citing Shared Resources
Researchers using the Fred & Pamela Buffett Cancer Center Shared Resources for publications, presentations or other scholarly outputs are required to acknowledge the use of them. Proper acknowledgment helps support the continued availability and funding of these cores.
Please acknowledge the Cancer Center Support Grant (P30 CA036727) in all publications and abstracts that have used these services.
Suggested language: Research reported in this publication was supported by the Flow Cytometry Shared Resource (RRID:SCR_017736) and the National Cancer Institute under award number P30 CA036727. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute.
Director
Craig L. Semerad, PhD
Director, Flow Cytometry Shared Resource
Durham Research Center 1042 | Send Email | 402-559-6299