The goal of this core is to provide sophisticated fluorescence-activated cell sorting and analytical services at reasonable hourly rates. To achieve these goals, the facility is operated to provide access to both clinical labs and basic science investigators.
As a clinically certified facility, the Flow Cytometry Core Facility performs clinical analyses of patient samples on a regular basis. In addition, five flow cytometers, including a 4-color FACSCalibur, a 10-color FACSCanto, a 15-color LSR II, a 16-color LSR Fortessa and a 16-color BD FACS Aria SORP High Performance cell sorter, are 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.
Laser excitation wavelengths (2):
Blue 488; Red 635nm
Fluorescence detection (4):
BD FACSCanto (10-color)
Laser excitation wavelengths (3):
Blue 488; Red 635nm; Violet 405nm
Fluorescence detection (10):
BD FACSAria SORP
Laser excitation wavelengths (4):
Blue 488; Red 635; Violet 405nm; Green 532nm
Fluorescence detection (16):
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.
Special Order BD LSR II
Laser excitation wavelengths (4):
Blue 488; Red 635; Green 532; Violet 405nm
Fluorescence detection (15):
BD LSR Fortessa
Laser excitation wavelengths (4):
Blue 488; Red 640; Green 532 nm; Violet 405nm
Fluorescence detection (16):
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.
Click on the above link to go to the Penn State Fluorofinder site.
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 LABLINK 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 from the DIVA workstations to your laboratory for archival. The server space (\\hersheymed.net\Files\research\corefacility\Results\Flow Cytometry) is to be used only as a dropbox for later transfer to your own archival system. 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. The DIVA workstations will only retain up to 3 months of your work in the DIVA database and Export folders. The research server is not a secure space to archive data so again, the user is responsible to archive flow cytometry data, not the core lab.
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 2 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.
Summary of Guidelines for the Safe Use of the Flow Cytometry Facility
The Flow Cytometry Facility operates as a Biosafety Level 2 laboratory. These guidelines are reviewed and revised if necessary each year and each facility user is responsible to review them on a regular basis to insure they are in compliance. Occasionally, some cell sorting experiments may operate in the cell sorter room under BSL-2+ conditions. This will be determined by the Flow Core Staff.
1) Admission to the laboratory is restricted to authorized personnel when work with human samples or a known infectious agent is in progress.
2) All cells must be assigned appropriate BSL levels for research use by the Biological Safety and Recombinant DNA Committee (Ralph Keil, Ph.D., Chair) before they are allowed to be used in the Flow Cytometry Core. Additional information can be found on the Safety Infonet website.
3) No BSL2+ samples are allowed to be run on the analyzers.
4) Before any cell sorting experiment can be scheduled, the Cell Sorting Experiment Form must be completed, signed and presented to the operator. This allows time for appropriate evaluation of biohazards under high speed sorting conditions. This must be done at least a week before the proposed experiment.
5) Some unfixed, primary human and primate cells and other BSL2 cells to be used on the FACSAria High Speed Cell Sorter may be designated as BSL2+ by the Flow Biosafety Committee. In these cases, strict adherence to the BSL2+ SOP will be required (located in the Flow Core). These same cells MAY be used under BSL2 conditions on the Cell Sorter if they are fixed according to accepted methods of inactivating all biohazards. (See note on Special Requirement for Sorting Primary Human Cells below.)
6) Whenever possible, samples of human or primate origin to be run on any cytometer should be fixed according to a process known and documented to inactivate HIV and other biohazards.
7) All procedures must be performed carefully to minimize the creation of aerosols.
8) All samples and time used on the instruments must be logged into the facility in Proteus. Samples that have been fixed for analyzing or sorting on the Cell Sorter may be monitored and recorded to insure that the fixation is adequate.
9) Work surfaces are to be decontaminated with 10% household bleach (FACS Clean) after any spillage of sample or hazardous material and after each user completes his/her work.
10) Instrument decontamination shall be performed according to protocols posted on each instrument.
11) All spills, however small, must be recorded in the accident book and reported to the Lab Manager.
12) All liquid and solid waste and disposal is to be removed from the laboratory as biohazardous material by each user. This includes all nuclear staining reagents such as PI, EB, AO, etc.
13) Gloves must be worn whenever handling or running biohazardous samples on the cytometer.
14) Hands must be washed after handling biohazardous materials and before leaving the laboratory.
15) Laboratory coats must be worn in the work area. These coats should be removed before leaving the laboratory.
16) Neither syringes nor hypodermic needles should be used in the facility without the consent of the Laboratory Manager. Glass objects should only be used when absolutely necessary.
17) No eating, drinking, smoking or applying cosmetics in the work area.
18) No mouth pipetting.
19) No animals are permitted within the laboratory.
20) All biohazard samples must be transported to and from the Flow Core in double sealed containers per College of Medicine safety policy.
In addition to the above guidelines and because this facility uses Class 3a through Class 4 lasers, the following Laser Safety guidelines shall also be observed:
- Access to the laboratory will be limited to only those persons necessary to the running of experiments when any laser is operating in the UV mode or when it is necessary to operate a laser without protective shielding.
- In such cases, the appropriate protective goggles or glasses will be made available and must be worn by those in the work area.
- Only qualified personnel shall operate or adjust any laser.
In the event of a potential retrovirus exposure
This item refers to exposure of skin or mucous membranes to infectious materials, and is modeled after the NIH program, "3 Emergency Steps to Take in the Event of a Potential HIV Exposure". Remember that the risk of infection is very dependent upon the titer of virus and the route of exposure. Thus the risk of infection by contact of intact skin with infectious body fluids is probably truly zero, and although the risk of infection by contact of mucous membranes or non-intact skin with infectious body fluids may be extremely small, it is likely to be no-zero.
The 3 emergency steps are:
- Immediately initiate first aid at the work site.
- Contaminated skin should be meticulously cleaned for 10 minutes using a povidone iodine solution (such as Betadine) and copious amounts of water.
- Contaminated eyes and mucous membranes should be irrigated for 5 minutes using water.
- Notify your supervisor, if he or she is immediately available. More importantly, go on to step 3.
- Report to the hospital Emergency Room to activate the invasive accident protocol (needlestick protocol), which will include evaluation, counseling, and provision of antiretroviral treatment if deemed appropriate. Do this immediately (i.e. within the half-hour).
General Sample Requirements
TUBES: Cells must be brought in Falcon 12x75mm polystyrene test tubes (Falcon # 2008) for running on any of the analyzers. Other tubes of varying sizes may be used on the cell sorter. Collection tubes for cell sorting should be polypropylene.
VOLUMES and CONCENTRATIONS: Minimum sample volumes per tube are .5 ml. Cell concentrations should be .5 - 1.0 X 106 cells per ml.
FLUOROCHROME SELECTION: Please review the capabilities of the instruments before selecting your fluorochromes and dyes to ensure that the instruments can excite and detect the dyes of choice. For multicolor experiments, it is imperative that you consult fluorochrome guidelines or personnel in the facility for best possible outcomes.
COMPENSATION CONTROLS: Bring positive and negative controls. A negative control is an unstained sample of cells in PBS or fixative or a sample of cells stained only with the secondary ab if this is the system you are using. If you have samples stained with more than one fluorochrome per test tube, bring in a sample stained with each fluorochrome individually. This is for compensation purposes. If you do not have enough positive stained cells for each fluorochrome, you may use antibody capture beads. To make conclusions about your experiment, the proper controls are critical. If you have any questions as to what the appropriate controls are, do not hesitate to call facility personnel at ext. 6908.
General Cell Staining Protocols
Cell Surface Marker/Antigen Staining
For each control and test sample, prepare a single cell suspension of 1X106 cells in cold PBS. Wash cells 1X in at least 1 ml of PBS with 2% FBS/media, centrifuge cells and aspirate supernatant.
Resuspend cells in a final volume of 50ul containing the primary antibody at the correct titer and PBS with 2% FBS/media. Reference manufacturer's guidelines but in general, incubate on ice or at room temp. for 20-30 minutes.
Wash cells 2X in PBS with 2% FBS/media. Centrifuge and aspirate supernatant.
For a directly conjugated antibody go to the next step. For indirect labeling, resuspend cell pellet in a final volume of 50ul containing the Secondary antibody and PBS with 2% FBS/media.
Resuspend cells at a final concentration of 1-2X106 cells/ml in at least 0.4ml fresh 1-2% formaldehyde/PBS. Provide the proper controls and analyze by flow cytometry.
Intracellular Staining- there are also good commercial kits available
- PI Staining of ETOH fixed cells for DNA Content, Cell Cycle and Apoptosis (Easiest Method)
- PI Staining of Nuclei for DNA Content and Cell Cycle (Best Resolution)
There are good commercial kits available. We recommend using kits with Annexin V PE and 7AAD as the best fluorochrome combination.
Current Protocols in Cytometry is available in the Flow Core upon request.
For information on other protocols, visit some of our other links or consult with resources in the Flow Core Facility.
Cell Sorting Requirements
Cells should be concentrated to at least 5 million cells/ml. If you want to sort lots of cells in a short amount of time, you may concentrate your cells even more (but the more concentrated your cells are, the more likely they might form clumps). Remember to calculate how many total cells you will need to analyze in order to sort the required number of target cells you want to have after the sort experiment. In general, lymphocyte sized cells can be run at rates close to 30K/sec. and cell lines can be run around 12K/sec.
You will need to bring cell media in the tubes or trays into which you want to sort cells. Tubes should be polypropylene to avoid static charges and can vary in size. Media should fill about 1/3 of the tube space before collection begins.
It is required that you filter your cells before sorting. Clogs will be a problem to any sorting experiment and increase biohazard risks. 35 or 40um filter cap tubes are available from BD Biosciences for this purpose.
Also, you must read and adhere to the SAFETY GUIDELINES before sorting unfixed cells and you must fill out and have the P.I. sign the CELL SORTING EXPERIMENT form before you can schedule any sorting experiment. This form must be given to the core personnel for review ONE WEEK prior to scheduling your sorting experiment.
If, after our review of the CELL SORTING EXPERIMENT form, it is determined that your experiment is at the BSL2+ level, you will be required to read and adhere to the Flow Cytometry BLS2+ SOP for cell sorting which is available in the cell sorter room.
Special Requirement for Sorting Primary Human Cells
Any request to sort live, primary human cells in the Flow Cytometry Core will require a review of the samples in question by the Flow Core Director based on the information on the CELL SORTING EXPERIMENT form and possible consultation with the PI involved. (This does not necessarily apply to human cells that are from an established published cell line.)
Several outcomes of this evaluation are possible. Some high risk populations may be deemed too high-risk to sort in our facility. Other populations may be sorted under BSL2 conditions, others under BSL2+ conditions and still others only after the donors are tested for HIV antibody and Hepatitis C and B Core antibody.
If donor testing is required, you must contact the Clinical Trials Coordinator for more information on this process. Once the tests are completed, a copy of the test report without patient ID will be signed by the requesting PI and forwarded to the Flow Cytometry Facility before the cell sorting experiment is performed.
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 to 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.
Click the link above to access the Genologics Lablink site.
Introductions to Flow Cytometry
- Video Introduction to Flow Cytometry
- Other Flow Cytometry Tutorials
- Free Online Book: "Flow Cytometry - A Basic Introduction" by Michael Ormerod
Fluorochrome Choices and Controls
- Core Facility Canto 10 Research Fluorochrome Chart
- Core Facility Fortessa Fluorochrome Chart
- Core Facility LSR II Fluorochrome Chart
- Core Facility FACSAria Fluorochrome Chart
- Core Facility Instrument Configurations
- BD Biosciences Multicolor Flow Tools (Spectral Viewer, Panel Designer, Fluorochrome Specifications)
- Multicolor Fluorochrome Guide
- Fluorochrome Relative Brightness
Cytoplasmic Staining Resources
- Current Protocols in Cytometry (see lab manager)
- Purdue University Cytometry (Very Comprehensive Resource)
- International Society for the Advancement of Cytometry
- Catalog of Free Flow Cytometry Software
- BD Biosciences (FACSCalibur, FACSCanto, LSR II, FACSAria)
- Molecular Probes (Invitrogen)
Other Related Resources
Use of Facility During Off Hours
Because of the nature of certain research projects it is sometimes necessary that accommodations be made for the processing of samples through the flow cytometers in the absence of the Flow Cytometry Core personnel who normally set up the instruments and assist the users for data acquisition. The Flow Cytometry Core has a Specialist and a trained backup Research Associate to cover almost all of the normal business hours for research use. However, there are rare occasions when neither person will be available to assist users even during regular business hours. For this reason, it is necessary to consult the Scheduling Calendar well in advance to ensure that your project can be scheduled accordingly.
For times other than regular business hours (i.e. evenings or weekends), certain approved users will also be able to use the flow cytometers upon prior arrangement with the Core personnel. There is a key pad lock on the door but the code will be made available for these users. These users will be completely "on their own" in starting up the instrumentation and shutting it down properly.
In the event that problems would arise, the operator has the option of calling the Lab Manager, Nate Sheaffer, at (717) 571-6231 (cell phone). If he can not be reached or the problem can not be solved over the phone, it will be necessary to stop the processing of samples at that point and wait until Core personnel return to the Lab. These special arrangements need to be scheduled ahead of time with the approval of the Core personnel.
To be approved for such "off hours" data acquisition, please see one of the Core personnel to complete the necessary training checklist for the particular instrument that will be used. For any occasion to process samples when Core personnel are not in the lab, only the Research FACSCalibur, FACSCanto II, LSR II and LSR Fortessa are available for research use. There are special arrangements for the other instruments to be made available for the processing of clinical specimens.
The FACSAria Cell Sorter will not be available for use in the absence of Core personnel at any time.
For information on fees please contact core facility personnel at (717) 531-6908.
Some recent publications which used data generated in the Flow Cytometry Core are listed below:
Distinct and Synergistic Roles of Fc¿RIIB Deficiency and 129 Strain-Derived SLAM Family Proteins in the Development of Spontaneous Germinal Centers and Autoimmunity. J Autoimmun (2015), http://dx.doi.ord/10.1016/j.jaut.2015.06.011. Soni C, Domeier PP, Wong EB, Shwetank, Khan TN, Elias MJ, Schell SL, Lukacher AE, Cooper TK, Rahman ZSM. article
Purification of Dendritic Cell and Macrophage Subsets from the Normal Mouse Small Intestine. J of Immunological Meth 421 (2015) 1-13. Koscso B, Gowda K, Schell TD, Bogunovic M. article
Peptide-MHC-I from Endogenous Antigen Outnumber Those from Exogenous Antigen, Irrespective of APC Phenotype or Activation. PloS Pathog. 2015 Jun; 11(6): e1004941. Sei JJ, Haskett S, Kaminsky LW, Lin E, Truckenmiller ME, Bellone CJ, Buller RM, Norbury CC. article
Macrophage-Derived TNF-a Mediates Diabetic Renal Injury. Accepted for Publication in Kidney International on June 10, 2015. PMID: 26061548. Awad AS, You H, Gao T, Cooper TK, Nedospasov SA, Vacher J, Wilkinson PF, Farrell FX, Reeves WB. article
B Cell-Intrinsic CD84 and Ly108 Maintain Germincal Center B Cell Tolerance. J Immunol 2015 Mar 23 pii. 1403023. Wong EB, Soni C, Chan AY, Domeier PP, Shwetank, Abraham T, Limaye N, Khan TN, Elias MJ, Chodisetti SB, Wakeland EK, and Rahman ZS. article
Protection from Tumor Recurrence Following Adoptive Immunotherapy Varies with Host Conditioning Regimen Despite Initial Regression of Autochthonous Murine Brain Tumors. Cancer Immunol Immunother 2015 Mar: 64(3): 325-336.1007/S00262-014-1635-7. Epub 2014 Nov 19. Cozza EM, Cooper TK, Budgeon LR, Christensen ND, and Schell TD. article
A Novel Role for the Mono-ADP-Ribosyltransferase PARP14/ARTD8 in Promoting Homologous Recombination and Protecting Against Replication Stress. Nucleic Acids Res. 2015 Mar 31; 43(6): 3143-3153. Nicolae CM, Aho ER, Choe KN, Constantin D, Hu HJ, Lee D, Myung K, Moldovan GL. article
Prolonged Antigen Presentation following an Acute Virus Infection Requires Direct and Then Cross-Presentation. Journal of Immun 2014 Oct 15 vol. 193 no. 8 4169-4177. Heipertz EL, Davies ML, Lin E and Norbury CC. article
Randomized controlled trial of oral glutathione supplementation on body stores of glutathione. Eur J Nutr. 2015 Mar;54(2):251-63. doi: 10.1007/s00394-014-0706-z. Epub 2014 May 5. Richie JP Jr, Nichenametla S, Neidig W, Calcagnotto A, Haley JS, Schell TD, and Muscat JE. article
B Cell-Intrinsic TLR7 Signaling is Essential for the Development of Spontaneous Germinal Centers. J Immunol 2014 Nov 1;193(9):4400-14. doi: 10.4049/jimmunol.1401720. Soni C, Wong EB, Domeier PP, Khan TN, Satoh T, Akira S, and Rahman ZSM. article
Targeted Disruption of a Ring-Infected Erythrocyte Surface Antigen (RESA)-like Export Protein Gene in Plasmodium falciparum Confers Stable Chondroitin 4-Sulfate Ctoadherence Capacity. ohJ Biolog Chem. 2014 Dec 5; 289, 34408-34421. Goel S, Muthusamy A, Miao J, Cui L, Salanti A, Winzeler EA, Gowda DC. article
Crosstalk between Muscularis Macrophages and Enteric Neurons Regulates Gastrointestinal Motility. Cell. 2014 July 17; 158: 300-313. Muller PA, Koscso B, Rajani GM, Stevanovic K, Berres ML, Hashimoto D, Mortha A, Leboeuf M, Li XM, Mucida D, Stanley ER, Dahan S, Margolis KG, Gershon MD, Merad M, and Bogunovic M. article
Whole-body Irradiation increases the Magnitude and Persistence of Adoptively Transferred T-cells Associated with Tumor Regression in a Mouse Model of Prostate Cancer. Cancer Immunol FRes 2014 Aug;2(8):777-88. doi: 10.1158/2326-6066.CIR-13-0164. Epub 2014 May 6. Ward-kavanagh LK, Zhu J, Cooper TK, and Schell TD. article
Tumour Cell Heterogeneity Maintained by Cooperating Subclones in Wnt-Driven Mammary Cancers. Nature. 2014 April 3; 508:113-117. Cleary, AS, Leonard, TL, Gestl, Shelley, Gunther, EJ. article
MyD88-Dependent Immunity to a Natural Model of Vaccinia Virus Infection Does Not Involve Toll-Like Receptor 2. J Virol. 2014 March vol 88 no. 6 3557-3567. Davies, ML, Sei JJ, Siciliano NA, Ren-Huan X, Roscoe R, Sigal LJ, Eisenlohr LC, and Norbury CC. article
Extracts of Devil's Club (Oplopanax horridus) Exert Therapeutic Efficacy in Experimental Models of Acute Myeloid Leukemia. Phytoth Res. 2014 February 23: 1308-1314. McGill CM, Alba-Rodriguez EJ, Li S, Benson CJ, Ondrasik RM, Fisher LN, Claxton DF and Barth BM. article
Glucocorticoid Inhibition of Activation-Induced Cytidine Deaminase Expression in Human B Lymphocytes. Mol and Cell Endocrinology Vol 382 Issue 2, 15 Feb 2014; pp.881-887. Benko AL, Olsen NJ, Kovacs WJ. article
CD36 Contributes to Malaria Parasite-Induced Pro-Inflammatory Cytokine Production and NK and T Cell Activation by Dendritic Cells. PloS One. 2013; 8(10): e77604. Gowda NM, Wu X, Kumar S, Febbraio M, Gowda DC. article
Macrophages Directly Mediate Diabetic Renal Injury. Am J Physiol Renal Physiol. 2103 Dec; 305(12):F1719-27. PMID: 24173355. You H, Gao T, Cooper TK, Reeves WB, Awad AS. article
Rapid Decrease in CD8+ T Cells Following Treatment of Mice With Exogenous Corticoserone. Bios 2013 Sept; 84(3):148-157. Bahr EC and Yorty JL. article
Protective Role of Small Pigment Epithelium-Derived Factor (PEDF) Peptide in Diabetic Renal Injury. Am J Physiol Renal Physiol. 2013 Sep 15; 305(6): F891-900. PMCID:PMC3761290. Awad AS, Gao T, Gvritishvili A, You H, Liu Y, Cooper TK, Reeves WB, Tombran-Tink J. article
PhotoImmunoNanoTherapy Reveals an Anticancer Role for Sphingosine Kinase 2 and Dihydrosphingosine-1-Phosphate. ACS Nano. 2013 Mar 26; 7(3):2132-44. Barth BM, Shanmugavelandy SS, Kaiser JM, McGovern C, Altinoglu EI, Haakenson JK, Hengst JA, Gilius EL, Knupp SA, Fox TE, Smith JP, Ritty TM, Adair JH, Kester M. article
Short-term Corticosterone Treatment Decreases the Early CD8+ T Cell Response to Simian Virus 40 Tumor Antigen but has no Impact on the Late CD8+ T Cell Response. Brain Behav Immun. 2013 Feb; 28:139-48. doi: 10.1016/j.bbi.2012.11.005. Yorty JL. article
In vitro growth inhibition of human cancer cells by novel honokiol analogs. Bioorg Med Chem. 2012 May 15; 20(10):3202-11. Lin JM, Prakasha Gowda AS, Sharma AK, Amin S. article
Systemic isotretinoin therapy normalizes exaggerated TLR-2-mediated innate immune responses in acne patients. J Invest Dermatol. 2012 Sep; 132(9):2198-205. Dispenza MC, Wolpert EB, Gilliland KL, Dai JP, Cong Z, Nelson AM, Thiboutot DM. article
Function of glycoprotein E of herpes simplex virus requires coordinated assembly of three tegument proteins on its cytoplasmic tail. Proc Natl Acad Sci U S A. 2012 Nov 27; 109(48):19798-803. Han J, Chadha P, Starkey JL, Wills JW. article
Modification of a Tumor Antigen Determinant To Improve Peptide/MHC Stability Is Associated with Increased Immunogenicity and Cross-Priming a Larger Fraction of CD8+ T Cells. J Immunol. 2012 Dec 15; 189(12):5549-60. Watson AM, Mylin LM, Thompson MM, Schell TD. article
CD36 Modulates Proinflammatory Cytokine Responses to Plasmodium Falciparum Glcoslphosphatidylinositols and Merozoites by Dendritic Cells. Parasite Immunol. 2012 jul; 34(7):372-82. doi: 10.1111;j.1365-3024.2012.01367.x. Kumar S, Gowda NM, Wu X, Gowda RN, Gowda DC. article
TLR9 and MyD88 are Crucial for the Development of Protective Immunity to Malaria. J Immunol. 2012 May 15; 188(10):503-85. doi: 10.4049/jimmunol. 1102143. Gowda NM, Wu X, Gowda DC. 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. J Immunol. 2011 Jan 1; 186(1):183-94. Hunzeker JT, Elftman MD, Mellinger JC, Princiotta MF, Bonneau RH, Truckenmiller ME, Norbury CC. article
Plasmodium falciparum: differential merozoite dose requirements for maximal production of various inflammatory cytokines. Exp Parasitol. 2011 Jan; 127(1):202-7. Wu X, Gowda NM, Gowda DC. article
Targeted indocyanine-green-loaded calcium phosphosilicate nanoparticles for in vivo photodynamic therapy of leukemia. ACS Nano. 2011 Jul 26; 5(7):5325-37. Barth BM, I Altinoglu E, Shanmugavelandy SS, Kaiser JM, Crespo-Gonzalez D, DiVittore NA, McGovern C, Goff TM, Keasey NR, Adair JH, Loughran TP, Claxton DF, Kester M. article
Overexpression of the dynein light chain km23-1 in human ovarian carcinoma cells inhibits tumor formation in vivo and causes mitotic delay at prometaphase/metaphase. Int J Cancer. 2011 Aug 1; 129(3):553-64. Pulipati NR, Jin Q, Liu X, Sun B, Pandey MK, Huber JP, Ding W, Mulder KM. article
Therapeutic efficacy of FTY720 in a rat model of natural killer cell leukemia (2011). Blood. 2011 September 8; 118(10): 2793–2800. 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
Synthesis and biological evaluation of a novel class of isatin analogs as dual inhibitors of tubulin polymerization and Akt pathway. Bioorg Med Chem. 2011 Oct 15; 19(20):6006-14. Krishnegowda G, Prakasha Gowda AS, Tagaram HR, Carroll KF, Irby RB, Sharma AK, Amin S. article
CD11b+, Ly6G+ cells produce type I interferon and exhibit tissue protective properties following peripheral virus infection. PLoS Pathog. 2011 Nov; 7(11):e1002374. Fischer MA, Davies ML, Reider IE, Heipertz EL, Epler MR, Sei JJ, Ingersoll MA, Rooijen NV, Randolph GJ, Norbury CC. article
Enhanced osteoclastic resorption and responsiveness to mechanical load in gap junction deficient bone. PLoS One. 2011; 6(8):e23516. Zhang Y, Paul EM, Sathyendra V, Davison A, Sharkey N, Bronson S, Srinivasan S, Gross TS, Donahue HJ. article
Renal dendritic cells ameliorate nephrotoxic acute kidney injury. J Am Soc Nephrol. 2010 Jan; 21(1):53-63. Tadagavadi RK, Reeves WB. article
FOXO3a elicits a pro-apoptotic transcription program and cellular response to human lung carcinogen nicotine-derived nitrosaminoketone (NNK). Lung Cancer. 2010 Jan; 67(1):37-47. Blake DC, Mikse OR, Freeman WM, Herzog CR. article
FoxP3aand Bcl-xL cooperatively promote regulatory T cell persistence and prevention of arthritis development (2010). Arthritis Res Ther. 2010;12(2):R66. doi: 10.1186/ar2983. Epub 2010 Apr 12. Rizwanul Haque, Fengyang Lei, Xiaofang Xiong, Yuzhang Wu and Jianxun Song article
Stress-induced glucocorticoids at the earliest stages of herpes simplex virus-1 infection suppress subsequent antiviral immunity, implicating impaired dendritic cell function. J Immunol. 2010 Feb 15; 184(4):1867-75. Elftman MD, Hunzeker JT, Mellinger JC, Bonneau RH, Norbury CC, Truckenmiller ME. article
Derivation of murine induced pluripotent stem cells (iPS) and assessment of their differentiation toward osteogenic lineage. J Cell Biochem. 2010 Mar 1; 109(4):643-52. Li F, Bronson S, Niyibizi C. article
Dendritic cell migration limits the duration of CD8+ T-cell priming to peripheral viral antigen. J Virol. 2010 Apr; 84(7):3586-94. Schell AM, Granger EL, Koczot F, Fischer MA, Norbury CC. article
A non-coding cationic lipid DNA complex produces lasting anti-leukemic effects. Cancer Biol Ther. 2010 Sep; 10(6):625-31. Keasey N, Herse Z, Chang S, Liggitt DH, Lay M, Fairman J, Claxton DF. article
FOXO3 encodes a carcinogen-activated transcription factor frequently deleted in early-stage lung adenocarcinoma. Cancer Res. 2010 Aug 1; 70(15):6205-15. Mikse OR, Blake DC, Jones NR, Sun YW, Amin S, Gallagher CJ, Lazarus P, Weisz J, Herzog CR. article
Direct presentation regulates the magnitude of the CD8+ T cell response to cell-associated antigen through prolonged T cell proliferation. J Immunol. 2010 Sep 1; 185(5):2763-72. Tatum AM, Watson AM, Schell TD. article
Endogenous IL-10 attenuates cisplatin nephrotoxicity: role of dendritic cells. J Immunol. 2010 Oct 15; 185(8):4904-11. Tadagavadi RK, Reeves WB. article
Read-through Activation of Transcription in a Cellular Genomic Context (2010). PLoS One. 2010; 5(12): e15704. Li Shen, David J. Spector article
PRAS40 Regulates Protein Synthesis and Cell Cycle in C2C12 Myoblasts (2010). Mol Med. 2010 Sep-Oct; 16(9-10): 359–371. Abid A Kazi and Charles H Lang article
Primary cell lines: false representation or model system? a comparison of four human colorectal tumors and their coordinately established cell lines (2010). Int J Clin Exp Med. 2010; 3(1): 69–83. Published online 2010 February 22. 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
Regulation of natural killer T-cell development by deubiquitinase CYLD (2010) EMBO J. 2010 May 5; 29(9): 1600–1612. Andrew J Lee1, Xiaofei Zhou, Mikyoung Chang, John Hunzeker, Robert H Bonneau, Dapeng Zhou and Shao-Cong Sun article
Targeting of survivin by nanoliposomal ceramide induces complete remission in a rat model of NK-LGL leukemia. Blood. 2010 Nov 18; 116(20):4192-201. Liu X, Ryland L, Yang J, Liao A, Aliaga C, Watts R, Tan SF, Kaiser J, Shanmugavelandy SS, Rogers A, Loughran K, Petersen B, Yuen J, Meng F, Baab KT, Jarbadan NR, Broeg K, Zhang R, Liao J, Sayers TJ, Kester M, Loughran TP. article
Viral sequestration of antigen subverts cross presentation to CD8(+) T cells. PLoS Pathog. 2009 May; 5(5):e1000457. Tewalt EF, Grant JM, Granger EL, Palmer DC, Heuss ND, Gregerson DS, Restifo NP, Norbury CC. article
Mechanisms of growth inhibition in human papillomavirus positive and negative cervical cancer cells by the chloromethyl ketone protease inhibitor, succinyl-alanine-alanine-proline-phenylalanine chloromethyl ketone. J Pharmacol Exp Ther. 2009 Jul; 330(1):359-66. Duncan KJ, Eckert KA, Clawson GA. article
Src activity alters alpha3 integrin expression in colon tumor cells. Clin Exp Metastasis. 2009; 26(2):77-87. Kline CL, Olson TL, Irby RB. article
Cooperative role of the MHR and the CA dimerization helix in the maturation of the functional retrovirus capsid. Virology. 2008 Jun 20; 376(1):191-8. Lokhandwala PM, Nguyen TL, Bowzard JB, Craven RC. article
If you have any other articles which used data from the Flow Cytometry Facility, please contact Nate Sheaffer at email@example.com.
Acknowledging the Core
Citation for Publications
Please cite the Penn State Hershey Flow Cytometry Core Facility and list the names of the instruments that were utilized either in the Materials and Methods section or the Acknowledgements section of your article.
Materials and Methods:
"Flow cytometric data were collected using an LSR II (Becton Dickinson) instrument in the Penn State Hershey Flow Cytometry Core Facility.
"Live cells were sorted by flow cytometry under BSL-2 conditions using a FACSAria SORP (Becton Dickinson) instrument in the Penn State Hershey Flow Cytometry Core Facility."
"We thank Nate Sheaffer and Joseph Bednarzyk from the Penn State Hershey Flow Cytometry Core Facility for assistance with flow cytometry analysis and cell sorting."
Flow Cytometry Instrumentation:
BD FACSCalibur (BD Biosciences, San Jose, CA)
BD FACSCanto II (BD Biosciences, San Jose, CA)
BD LSR II (Special Order System), (BD Biosciences, San Jose, CA)
BD LSRFortessa (BD Biosciences, San Jose, CA)
BD FACSAria (Special Order Research Product), (BD Biosciences, San Jose, CA)
Todd Schell, PhD
Director, Flow Cytometry Core
Phone (717) 531-8169
Flow Cytometry Core Advisory Panel
- Bruce Stanley, Ph.D., Director of Research Resources
- Todd Schell, Ph.D., Director of Flow Cytometry Core
- Nate Sheaffer, Core Lab Manager, Flow Cytometry Specialist
- Christopher Norbury, Ph.D., Research Cytometry and Biosafety Advisor
- Jeff Sample, Ph.D., Biosafety Advisor
- Mike Bayerl, M.D., Clinical Cytometry Advisor
The daily operations as well as the review of clinical samples are the responsibility of:
| Nate Sheaffer, BA, ASCP Cyt |
Core Lab Manager
Phone (717) 531-6908
Joseph Bednarczyk, BS
| Jade Vogel, BS |
Phone (717) 531-6908