Flow Cytometry Core
The Flow Cytometry Core provides investigators with a resource for analytical and preparative studies of cells using flow cytometry. The Core offers FACS acquisition and analysis, staining for apoptosis and DNA cell cycle, cell sorting, and consultation for experimental design, interpretation and troubleshooting. Applications include immunophenotyping, intracellular staining, molecular and cellular probes such as Ca flux and GFP, CBA (Cytometric Bead Array), DNA cell cycle, TUNEL and Annexin V for apoptosis, sterile sorting for cloning and more.
New to the Flow Core? (see guide) or
download NewtoFlowCore.pdf
The Core is equipped with the following instruments:
- Becton-Dickinson LSRII capable of 12 color and 14 parameter acquisition with 5 lasers (Argon ion 488nm, HeCd UV 355nm, Violet 405nm, HeNe 639nm, and Green 532nm).
- Two Becton-Dickinson FACScans capable of 3 color and 5 parameter acquisition with an Argon ion laser (488nm).
- Becton-Dickinson Special Order BD FACSAria II high performance, fixed alignment cuvette cell sorter. The Special Order BD FACSAria II consists of 488nm, 640nm, 405nm, 561nm and 355nm lasers, providing 16 parameters.
- BioProtect III Walk In Clean Air and Containment Cabinet houses the FACSAria II. The BioProtect III is a specialty type Class II biological safety enclosure designed to offer personnel, product, and environmental protection from potentially aerosolized biological hazards.
- Invitrogen Countess™ automated cell counter.
The Core provides data analysis with the following software:
- FACSDiva (Becton-Dickinson)
- Cell Quest (Becton-Dickinson)
- Flow Jo (Tree Star)
- ModFit LT (Verity)
Analysis workstations and software training are available for researchers.
Please schedule appointments for analysis and sorting at 216-445-3798. The Flow Cytometry Core is located in NBb-40. The FACSAria II is located in NE3-224.
- Services
- Equipment
- Personnel
- Protocols
The Core offers many services for the analysis or isolation of cells following staining of cellular proteins or DNA with fluorochrome labeled reagents.
FACS applications include:
Consultation
All investigators are encouraged to meet with the Flow Cytometry Core technologists to discuss the experiment. This meeting is often useful in that the technical limitations of an experiment are realized and many potential problems can be averted before critical resources are used. The Core can also provide written protocols for staining, fixation and data analysis methods. The Core technologists are available for assistance with data retrieval, analysis and presentation.
Materials available for reference in the Flow Core lab include several textbooks on the basics of flow cytometry and the Current Protocols in Cytometry. In addition, the Core frequently organizes training seminars at different levels of expertise.
Appointments
Investigators must schedule appointments in advance for ALL flow cytometry. Currently 1-2 days notice is required; however, more notice is necessary for sorting. Appointments can be made by phone or e-mail. All experiments must be accompanied by appropriate controls.
BD FACSAria II
The Flow Cytometry Core has acquired a new high performance, fixed alignment cuvette cell sorter that makes sorting easier and more accessible. The Stem Cell Biology and Regenerative Medicine Department courteously funded the Special Order BD FACSAria II. The new sorter improves ease of use, flexibility, and aseptic capability making it the best choice for consistent results in sorting across a broad range of applications.
A BioProtect III Walk In Clean Air and Containment Cabinet houses the FACSAria II. The BioProtect III is a specialty type Class II biological safety enclosure designed to offer personnel, product, and environmental protection from potentially aerosolized biological hazards.
The Special Order BD FACSAria II consists of 488nm, 640nm, 405nm, 561nm and 355nm lasers, providing 16 parameters. The optical filters detect the following fluorophores:
(AriaII_config.pdf)
- Blue laser - FITC, PerCP-Cy5.5
- Red laser - APC-Cy7, Alexa 700, APC
- Yellow-Green laser - PE, PE-Cy5, PE-Cy7, PE-TxRd, mCherry and DsRed
- Violet laser - Pacific Blue, Qdot 585, Qdot 605, Qdot 655, AmCyan and Qdot 565
- UV laser - DAPI, Hoechst, Indo-1, Side Population Filter Set
Additional options included:
- 70-, 85-, 100- and 130-micron nozzle tips
- Two- and four-way sorting into microtubes, 12 x 75-mm tubes, and 15 mL tubes
- Accepts various sizes of sample input tube
- Automated Cell Deposition Unit (ACDU) allows for multi-well plate and slide sorting
- Cooling system for the sort collection device (tubes and multi-well plates)
- Aerosol Management for enhanced removal of aerosols
Countess
The Flow Cytometry Core is now offering the use of the Countess™ automated cell counter. The Countess™ provides fast, easy and accurate cell counting without using a hemocytometer, eliminating the tedium and subjectivity of manual cell counting. Automated counting frees up your time and minimizes subjective judgments that can lead to error. (Countess.pdf)
The Countess(TM) instrument:
- Counts live and dead cells
- Calculates percent viability
- Measures average cell size
- Calculates dilutions for downstream applications
- Uses just 10 microliters of sample
- Saves your data to a USB drive
BD FACScan
Scanning is achieved by using a Becton-Dickinson FACScan flow cytometer. The instrument has a highly sensitive laser used exclusively for quantitative analysis. The FACScan is equipped with three detectors to measure fluorochromes that excite at 488 nm. The FACScan is engineered with fixed optical, electronic and fluidic components that cannot be reconfigured.
(FACScan_config.pdf)
BD LSRII
Scanning is achieved by using the Becton-Dickinson LSRII flow cytometer. The instrument has highly sensitive lasers used exclusively for quantitative analysis. The LSRII is capable of 12 color and 14 parameter acquisition with 5 lasers (Argon ion 488nm, HeCd UV 355nm, Violet 405nm, HeNe 639nm, and Green 532nm).
(LSRII_config.pdf )
Clemencia Colmenares
- Clemencia Colmenares PhD
- Director/Staff
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Phone:(216)-445-6613
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Cathy Shemo
- Cathy Shemo BS
- Lead Research Technologist
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Phone:(216)-445-3798
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Sage O'Bryant
- Sage O'Bryant
- Research Technologist
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Phone:(216)-445-3798
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Bunny Cotleur
- Bunny Cotleur MS
- Scientific Consultant
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Phone:(216)-444-1164
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Moneen Morgan
- Moneen Morgan BS
- Research Technician
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Phone:(216) 445-3798
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