The Flow Cytometry Shared Resource provides researchers with ready access to the necessary expertise and state-of-the-art instrumentation they need to explore a wide range of research problems with flow cytometry.
We focus on addressing the needs of seasoned flow cytometry users as well as those just discovering the capabilities of flow cytometry technology. We provide monthly training for the instruments for new users. Our directors are also available to consult on experimental design and data analysis.
Focus Areas
Typical experiments handled by the Flow Cytometry Shared Resource facility are designed to analyze or to sort based on up to nine fluorescent parameters. Some representative examples of analysis experiments include the:
- Measurement of DNA content for cell cycle analysis
- Cell cycle-specific nuclear and cytoplasmic proteins
- Apoptosis markers
- Substituted deoxyuridine incorporation
- Stem cell side populations
- Reactive oxygen species
- Cell surface markers using labeled antibodies or substrates
- Cell viability
- Intracellular ion concentration or pH
- Membrane potential
Cell lysates can also be analyzed with bead substrate-based immunoassays. Some representative examples of sorting experiments include:
- Purification of cells transfected with fluorescent protein-tagged DNA inserts
- Isolation of cell populations expressing specific intracellular or membrane metabolic products
Services Provided
Flow cytometry is a powerful, versatile technology that allows researchers to simultaneously analyze individual cells in a multiparameter fashion. We offer sterile cell sorting, which allows researchers to identify distinct subpopulations in a mixed population of cells and isolate up to four populations concurrently. We also provide analysis instruments, training for use of the instruments, access to analysis software, assistance in assay design and data analytics.
Use of the Instruments
Institutional instruments available include:
- BD FACS Aria cell sorter (three lasers and 11 parameters)
- Sorting tips available: 70 micron, 100 micron and 130 micron
- Capable of single-cell sorting into 24 and 96 well plates
- Instrument is fitted with a special flow cell to provide better precision for measuring DNA content and side-populations
- BD FACS Calibur analyzer (two lasers and six parameters)*
- BD FACS Fortessa X-20 analyzer (five lasers and 20 parameters)*
- Beckman-Coulter Astrios EQ cell sorter (five lasers and twenty parameters)
- Jet-in-area optics
- Multiple sorting tip options
- Capable of single-cell sorting into 24 and 96 well plates
- High throughput 6-way sorting
- BD Accuri C6 analyzer (two lasers and six parameters)*
- BD FACS Canto II analyzer (three lasers and 10 parameters)*
- BD FACS Calibur analyzer (two lasers and six parameters)*
- In addition to the software available with each flow cytometer, FCS Express flow cytometer analysis software is available to analyze the multiparameter data to identify and characterize cell subpopulations. DNA histogram analysis software programs (ModFit and MultiCycle) are also available to address the needs of researchers requiring advanced cell cycle analysis.
*Instruments are available to use 24 hours a day, seven days a week. Users must receive appropriate instruction to become approved instrument operators (see below).
Hands-on training is provided by Dr. John Whitesides, -9465, [email protected] and Dr. James Wood, -4461, [email protected] for individuals who would like the ability to acquire their data on the FACS Calibur, BD Acurri C6, FACS Canto, and FACS Fortessa X20. Please contact either Dr. Whitesides or Dr. Wood directly to schedule training.
To schedule a monthly training session please see the contact below for specific equipment:
| Equipment | Training Contact |
|---|---|
| FACS Calibur
| Dr. James Wood
|
| FACS Fortessa X20 | Dr. John F. Whitesides -9465 [email protected] |
Program key: CPC = Cancer Prevention and Control Program; CRP = Clinical Investigation Program; TPR = Tumor Progression and Recurrence Program; CBB = Cancer Biology and Biochemistry Program
Birbrair A, Sattiraju A, Zhu D, Zulato G, Batista I, Nguyen VT, Messi ML, Solingapuram Sai KK, Marini FC, Delbono O, Mintz A. Novel peripherally derived neural-like stem cells as therapeutic carriers for treating glioblastomas. Stem Cells Transl Med. 2017 Feb;6(2):471-481. doi: 10.5966/sctm.2016-0007. Epub 2016 Sep 14. PMID: 28191774 http://onlinelibrary.wiley.com/doi/10.5966/sctm.2016-0007/
Wu K, Fukuda K, Xing F, Zhang Y, Sharma S, Liu Y, Chan MD, Zhou X, Qasem SA, Pochampally R, Mo YY, Watabe K. Roles of the cyclooxygenase 2 matrix metalloproteinase 1 pathway in brain metastasis of breast cancer. J. Biol. Chem. 2015;290(15):9842-54. PMC4392281.
Swanner J, Mims J, Carroll DL, Akman SA, Furdui CM, Torti SV, Singh RN. Differential cytotoxic and radiosensitizing effects of silver nanoparticles on triple-negative breast cancer and non-triple-negative breast cells. Int J Nanomedicine. 2015;10:3937-53. PMC4501353.
