Flow Cytometry
The Penn State Hershey Flow Cytometry Core Facility provides sophisticated fluorescence-activated cell sorting and analytical services at reasonable hourly rates. The facility is operated to provide access to both clinical labs and basic science investigators. |  |
As a clinically certified facility, the performs clinical analyses of patient samples on a regular basis. In addition, four flow cytometers are dedicated to research, including a 4-color FACSCalibur, an 8-color FACSCanto, a 15-color LSR II, and a 16-color FACSAria SORP high speed cell sorter, all available for use by investigators at the College of Medicine.
Please take a few minutes to look over the website pages to ensure your flow cytometry experiments are conducted using appropriate methods and techniques to produce the most accurate data in a cost efficient manner.
Flow Cytometry Basics
Flow cytometry is a means of identifying and measuring certain physical and chemical characteristics of cells or particles as they travel in suspension one by one past a sensing point. The flow cytometer is able to "look" at thousands of cells or particles per second and perform and record many simultaneous measurements for each cell or particle.
The flow cytometer consists of a light source, collection optics, electronics and a computer to translate signals to data. The light source of choice is usually a laser which emits coherent light at a specified wavelength. Scattered and emitted fluorescent light is collected by two lenses (one set in front of the light source and one set at right angles) and by a series of optics, beam splitters and filters, specific bands of fluorescence can be measured.
A flow cytometer can measure physical characteristics such as cell size, shape and internal complexity and, of course, any cell component or function that can be detected by a marker attached to a fluorescent compound can be examined. A number of these cell markers and measurements can be made for each cell and combined to give an informative summary of the characterization, identification and function of large populations of cells. So the applications of flow cytometry are numerous, and this has led to the widespread use of these instruments in the biological and medical fields. See Helpful Resources for more information and introductions to Flow Cytometry.
 Diagram courtesy of DakoCytomation | Some of the more common research applications include: immunology, cell cycle and cell growth, cell function and activation, cell differentiation, apoptosis, platelet activation, toxicology and Green Fluorescent protein detection. Some of the common clinical studies include: leukemia and lymphoma characterization, immune studies such as T and B cell subset determinations, stem cell content monitoring for transplant, nuclear ploidy and cell cycle determinations and reticulocyte counting. In addition to analyzing populations of cells and particles for information and data which can be stored electronically and displayed in the form of dot plots and histograms, some flow cytometers have the ability to physically sort out cells or particles of interest. These cells can be sorted out of a heterogeneous mixture into a very pure population for further studies. |
Scheduling for All Flow Cytometers
Scheduling time for sample acquisition or data analysis is done by calling the Core facility at Ext. 6908. Researchers are scheduled on a first come first served basis. A Scheduling Calendar has been made available for all users to view online to plan experiments. Please "bookmark" the link for future use. It is "view only" and will change often so be sure to refresh the site as you make your plans. Scheduling is best done a few days in advance to ensure that you have sufficient time and that you will be able run your samples within a reasonable time after they have been prepared. If you are doing a time course study, it is best to check with the facility well in advance to ensure that someone will be there on the days you need to have access to the cytometers. Occasionally a request for live cell analysis will take priority over a previously scheduled time slot using fixed cells.
Requests for cell sorting must be made at least a week in advance to ensure assistance will be available and that all biosafety concerns can be addressed.
Rates for Flow Cytometer Use (Updated March 1, 2010)
Standard Sample Acquisition (Researcher provides operator) $50/hour
Standard Data Analysis (Researcher provides operator) $20/hour
Assisted Sample Acquisition or Data Analysis (Core provides operator) $50/hour
Standard Sorting on FACSAria (Researcher provides assistant) $85/hour
Sorting Setup Charge $100/sort
Special Sorter Setup Charge (i.e. nozzle change, plates) $150/sort
Minimum Charge for Sorting Use $185
NOTE: "No Shows" and cancellations less than 8 hours in advance may be charged for scheduled time.
General Procedures for the Core
The Flow Cytometry Core Facility serves both the Hospital and the Researchers of the College of Medicine. There are three instruments (FACSCanto and 2 FACSCaliburs) dedicated to clinical use. Use of these machines by researchers are prohibited unless specifically directed to do so by the Core personnel. All other instruments, including the Cell Sorter, are for research use and must be scheduled in advance for use (see above).
Billing for usage is done monthly according to Department and Principal Investigator. We do not break down the billing by technician or project, but if the researcher needs this information, they can retrieve it upon request to the core facility. All users must fill out the Proteus web page information each time they use equipment.
The facility is designed to have the users operate the instruments when running samples and analyzing data. The Core personnel will train these users to become proficient over time in setting up their experiments, running samples and producing and archiving data resulting from the experiments.
There is a terabyte server available to all the researchers to use for transferring data to your laboratory for archival or further analysis. Users may also transfer data to other portable storage media if desired. In any case, the user is ultimately responsible for archiving any and all data generated.
Safety for all users and Core personnel is a priority concern. Since the Core facility is a shared laboratory and is designated as a BSL II facility, there are certain guidelines that all users are responsible to be aware of and follow. This is a special concern for cell sorting and biosafety concerns and guidelines will be strictly adhered to. This policy is summarized in the Safety Guidelines. They include guidelines for the safe use of lasers as well.
BD FACSCaliburLaser excitation wavelengths (2): Blue 488; Red 635nm Fluorescence detection (4): 530, 585, >650; >670 |  |
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BD FACSCanto IILaser excitation wavelengths (3): Blue 488; Red 635nm; Violet 405nm Fluorescence detection (8): >Help in selecting appropriate fluorochromes for this Canto II Workstation: |  |
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BD FACSAria SORPLaser excitation wavelengths (4): Blue 488; Red 635; Violet 405nm; Green 532nm Fluorescence detection (16):
Workstation: Sorting Parameters: |  |
Users are expected to accompany their samples and assist in the monitoring of data acquisition and sorting experiments. The Core personnel will be responsible for all the set up and shut down procedures related to each experiment. For more information on the policies for using the FACSAria please see the Scheduling/Fees/General Procedures and the General Sample Requirements and Protocols. The cell sorting experiment report must be submitted prior to scheduling a sort on the FACSAria. ________________________________________________________
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Special Order BD LSR II Flow CytometerLaser excitation wavelengths (4): Blue 488; Red 635; Green 532; Violet 405nm Fluorescence detection (15):
> Help in selecting appropriate fluorochromes for this LSR II > Download BD's fluorochrome guide Workstation: PC based, DIVA Software |  |
Director
Bruce Stanley, PhD
Director, Research Resources
Phone (717) 531-5329
Email bas12@psu.edu
Flow Cytometry Core Advisory Panel
- Dr. Bruce Stanley, Director of Research Resources
- Nate Sheaffer, Laboratory Manager, Flow Cytometry Specialist
- Dr. Christopher Norbury
- Dr. Mike Bayerl
Lab Personnel
The daily operations as well as the review of clinical samples are the responsibility of:
| Nate Sheaffer, BA, ASCP Cyt Lab Manager & Flow Cytometry Specialist Room C3603 Phone (717) 531-6908 Email nas2@psu.edu |  |
Joseph Bednarczyk
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| David R. Stanford, Ph.D. Senior Research Support Associate Room C3603 Phone (717) 531-4699 Email drs19@psu.edu |  |
Some recent publications which used data generated in the Flow Cytometry Core are listed below
Direct Presentation Regulates the Magnitude of the CD8+ T Cell Response To Cell-Associated Antigen through Prolonged T Cell Proliferation (2010)
Angela M. Tatum1, Alan M. WatsonTodd D. Schell article
A Marked Reduction in Priming of Cytotoxic CD8 + T Cells Mediated by Stress-Induced Glucocorticoids Involves Multiple Deficiencies in Cross-Presentation by Dendritic Cells (2010)
E. Truckenmiller and Christopher C. Norbury
Mellinger, Michael F. Princiotta, Robert H. Bonneau, Mary
John T. Hunzeker, Michael D. Elftman, Jenniferdoi: article
Renal Dendritic Cells Ameliorate Nephrotoxic Acute Kidney Injury (2010)
Raghu K. Tadagavadi* and W. Brian Reeves article
Enhanced Osteoclastic Resorption and Responsiveness to Mechanical Load in Gap Junction Deficient Bone (2011)
Yue Zhang, Emmanuel M. Paul, Vikram Sathyendra, Andrew Davison, Neil Sharkey, Sarah Bronson, Sundar Srinivasan, Ted S. Gross, Henry J. Donahue article
PRAS40 Regulates Protein Synthesis and Cell Cycle in C2C12 Myoblasts (2010)
Abid A Kazi and Charles H Lang article
Therapeutic efficacy of FTY720 in a rat model of natural killer cell leukemia (2011)
Aijun Liao, Kathleen Broeg, Todd Fox, Su-Fern Tan, Rebecca Watters, Mithun Vinod Shah, Lucy Q. Zhang, Yongping Li, Lindsay Ryland, Jun Yang, Cesar Aliaga, Alden Dewey, Andrew Rogers, Kelly Loughran, Leah Hirsch, Nancy Ruth Jarbadan, Kendall Thomas Baab, Jason Liao, Hong-Gang Wang, Mark Kester, Dhimant Desai, Shantu Amin, Thomas P. Loughran, Jr. and Xin Liu article
A non-coding cationic lipid DNA complex produces lasting anti-leukemic effects (2010)
Nikki Keasey, Zachary Herse, Stella Chang, Denny H. Liggitt, Marla Lay, Jeffery Fairman and David F. Claxton article
Regulation of natural killer T-cell development by deubiquitinase CYLD (2010)
Andrew J Lee1, Xiaofei Zhou, Mikyoung Chang, John Hunzeker, Robert H Bonneau, Dapeng Zhou and Shao-Cong Sun article
Dendritic Cell Migration Limits the Duration of CD8 + T-Cell Priming to Peripheral Viral Antigen (2010)
Amanda M. Schell, Erica L. Granger, Frank Koczot, Matthew A. Fischer and Christopher C. Norbury article
FoxP3aand Bcl-xL cooperatively promote regulatory T cell persistence and prevention of arthritis development (2010)
Rizwanul Haque, Fengyang Lei, Xiaofang Xiong, Yuzhang Wu and Jianxun Song article
Endogenous IL-10 attenuates cisplatin nephrotoxicity: role of dendritic cells (2011)
Raghu Kempegowda Tadagavadi and William Brian Reeves article
CD11b+, Ly6G+ Cells Produce Type I Interferon andExhibit Tissue Protective Properties Following Peripheral Virus Infection (2011)
Matthew A. Fischer, Michael L. Davies, Irene E. Reider, Erica L. Heipertz, Melanie R. Epler, Janet J. Sei, Molly A. Ingersoll, Nico Van Rooijen, Gwendalyn J. Randolph, Christopher C. Norbury article
Primary cell lines: false representation or model system? a comparison of four human colorectal tumors and their coordinately established cell lines (2010)
Danielle M. Pastor, Lisa S. Poritz, Thomas L. Olson, Christina L. Kline, Leonard R. Harris III, Walter A. Koltun, Vernon M. Chinchilli, Rosalyn B. Irby article
Read-through Activation of Transcription in a Cellular Genomic Context (2010)
Li Shen, David J. Spector article
Systemic Isotretinoin Therapy Normalizes Exaggerated TLR-2-Mediated Innate Immune Responses in Acne Patients (2012)
Melanie C. Dispenza, Ellen B. Wolpert, Kathryn L. Gilliland, Jenny P. Dai, Zhaoyuan Cong,
Amanda M. Nelson and Diane M. Thiboutot article preview
Targeting of survivin by nanoliposomal ceramide induces complete remission in a rat model of NK-LGL leukemia (2010)
Xin Liu, Lindsay Ryland, Jun Yang, Aijun Liao, Cesar Aliaga, Rebecca Watts, Su-Fern Tan, James Kaiser, Sriram S. Shanmugavelandy, Andrew Rogers, Kathleen Loughran, Bailey Petersen, Jonathan Yuen, Fanxue Meng, Kendall Thomas Baab, Nancy Ruth Jarbadan, Kathleen Broeg, Ranran Zhang, Jason Liao, Thomas Joseph Sayers, Mark Kester and Thomas P. Loughran, Jr article
If you have any other articles which used data from the Flow Cytometry Facility, please contact Nate Sheaffer at nas2@psu.edu.