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. The FCSR critically enables studies at the single-cell level, and recent institutional investments in a new cell sorter/analyzer have broadened the range of cytometry applications, particularly in immuno-oncology. 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 18 fluorescent plus 6 light scatter 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
- Day GL, Bryan ML, Northrup SA, Lyles DS, Westcott MM, and JH Stewart 4th. Immune Effects of M51R Vesicular Stomatitis Virus Treatment of Carcinomatosis From Colon Cancer. J Surg Res. 2019 Aug 12;245:127-135. doi: 10.1016/j.jss.2019.07.032. [Epub ahead of print] PMID: 31415934.
- Deng C, Xie Y, Zhang C, Ouyang B, Chen H, Lv L, Yao J, Liang X, Zhang Y, Sun X, Deng C, Liu G. Urine-Derived Stem Cells Facilitate Endogenous Spermatogenesis Restoration of Busulfan-Induced Nonobstructive Azoospermic Mice by Paracrine Exosomes.Stem Cells Dev. 2019 Sep 9. doi: 10.1089/scd.2019.0026. [Epub ahead of print] PMID: 31311428
- Wajih N, Basu S, Ucer KB, Rigal F, Shakya A, Rahbar E, Vachharajani V, Guthold M, Gladwin MT, Smith LM, Kim-Shapiro DB. Erythrocytic bioactivation of nitrite and its potentiation by far-red light. Redox Biol. 2019 Jan;20:442-450. doi: 10.1016/j.redox.2018.11.001. Epub 2018 Nov 3. PMID: 30423533
- Phipps JP, Haas KM. An Adjuvant That Increases Protective Antibody Responses to Polysaccharide Antigens and Enables Recall Responses. J Infect Dis. 2019 Jan 7;219(2):323-334. doi: 10.1093/infdis/jiy506. PMID: 30289460
- Haro MA, Dyevoich AM, Phipps JP, Haas KM. Activation of B-1 Cells Promotes Tumor Cell Killing in the Peritoneal Cavity. Cancer Res. 2019 Jan 1;79(1):159-170. doi: 10.1158/0008-5472.CAN-18-0981. Epub 2018 Sep 17. PMID: 30224373
- Park S.H., Eber M.R., Taichman R.S., Shiozawa Y. Determining Competitive Potential of Bone Metastatic Cancer Cells in the Murine Hematopoietic Stem Cell Niche(2018) In: Turksen K. (eds) Stem Cell Niche. Methods in Molecular Biology, vol 2002: 141-150. Humana, New York, NY PMID: 30099699
- Gao W, Xu Y,Chen T,Du Z,Liu X, Hu Z, Wei D, Gao C, Zhang W, Li Q. Targeting oxidative pentose phosphate pathway prevents recurrence in mutant Kras colorectal carcinomas PLoS Biol 17(8): e3000425.https://doi.org/10.1371/journal.pbio.3000425 PMID: 31461438
- Tian W, Teng F, Gao J, Gao C, Liu G, Zhang Y, Yu S, Zhang W, Wang Y, Xue F. Estrogen and insulin synergistically promote endometrial cancer progression via crosstalk between their receptor signaling pathways. Cancer Biol Med. 2019 Feb;16(1):55-70. doi: 10.20892/j.issn.2095-3941.2018.0157. PMID: 31119046
- Bonomi M, Ahmed T, Addo S, Kooshki M, Palmieri D, Levine BJ, Ruiz J, Grant S, Petty WJ, Triozzi PL. . Circulating immune biomarkers as predictors of the response to pembrolizumab and weekly low dose carboplatin and paclitaxel in NSCLC and poor PS: An interim analysis. Oncol Lett. 2019 Jan;17(1):1349-1356. doi: 10.3892/ol.2018.9724. Epub 2018 Nov 19. PMID: 30655905
- Moreno NS, Liu J, Haas KM, Parker LL, Chakraborty C, Kron SJ, Hodges K, Miller LD, Langefeld C, Robinson PJ, Lelièvre SA, Vidi PA. The nuclear structural protein NuMA is a negative regulator of 53BP1 in DNA double-strand break repair. Nucleic Acids Res. 2019 Sep 12. pii: gkz802. doi: 10.1093/nar/gkz802. [Epub ahead of print] No abstract available. PMID: 31511892
- Alkhateeb T, Kumbhare A, Bah I, Youssef D, Yao ZQ, McCall CE, El Gazzar M. S100A9 maintains myeloid-derived suppressor cells in chronic sepsis by inducing miR-21 and miR-181b. Mol Immunol. 2019 Aug;112:72-81. doi: 10.1016/j.molimm.2019.04.019. Epub 2019 May 9. PMID: 31078