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Core FC

Core H redThe UAB Flow Cytometry Facility is comprised of two locations as a result of a consolidation between the flow cytometry cores of the Center for AIDS Research and the Comprehensive Arthritis, Musculoskeletal and Autoimmunity Center. These locations include BBRB (Bevill Biomedical Research Building, room 557) and Shelby (Shelby Biomedical Research Building, room 271). Our mission is to provide state-of-the-art cell sorting and analytical flow cytometry services to support research programs. The facility is made available to all Investigators at UAB and encourages collaborations and synergy.  Specific services and equipment at each location is listed below. Most services are provided at either location. Training, consultation, assisted- and self-serve equipment usage is available.

Locations

cfarcoreThe BBRB location provides the following equipment and services:

  • Newest Instrumentation
    • BD Accuri C6 Flow Cytometer
  • Cell sorting
    • BD FACS Aria
  • Cell analysis
    • BD LSR II GUAVA EasyCyte
  • Bioplex analysis
  • High throughput flow cytometry
    • Hypercyt autosampler
apcfcoreThe Shelby location provides the following equipment and services:

  • Cell sorting
    • BD FACS Aria
  • Cell analysis
    • BD LSR II
    • FACS Calibur
    • FACS Scan
  • AutoMacs
    • MACS Sorter

 

Scheduling


First Time Users:
Please contact Jena L. Reeves at jena@uab.edu to be set up as a user.

Once Ms. Reeves confirms that you have been set up, you will receive an email invitation with a link to the Priority Software online scheduling site. This link will provide you with instructions on how to set your password and log in to the scheduler. 

You now have Web Access to the BBRB Flow Cytometry Facility or the Shelby Flow Cytometry Facility. Your logon email address is:  blazerid@uab.edu

Questions:

If you have any questions or problems with the login, please contact Ms. Reeves by email or phone: 205-934-7753.

Contact the facilities for questions related to capabilties, training, equipment or scheduling.

Flow Cytometry Web Scheduling

Services


In 2010, the CAMAC (Comprehensive Arthritis, Musculoskeletal and Autoimmunity Center) and the CFAR (Center for AIDS Research) Flow Cytometry Cores officially established a partnership through a federal core consolidation grant.  This funding was provided by an ARRA supplemental award (3P30AI027767-22S1, PI: Dr. Michael Saag) to the UAB CFAR. These efforts lead to the purchase of a new 4-laser - 14 parameter FACSAria sorter to the CAMAC Flow Core and an upgrade of the existing CFAR Flow Core FACSAria II sorter. As a result of this consolidation, expanded equipment and services are now provided to all researchers at UAB and beyond. Please see what is available to support your studies below or contact us for more information.

accuri overview The BD Accuri C6 Cytometer system is equipped with a blue and red laser, two light scatter detectors, and four fluorescence detectors with optical filters optimized for the detection of FITC, PE, PerCP, and APC. A compact optical design, fixed alignment, and pre-optimized detector settings make the system easier to use. 

Location: BBRB 557

srv-hypercytautosampler
The HyperCytautosampler allows for high throughput flow cytometric analysis. Samples from 96- or 384 well plates are analyzed in a time resolved manner. The HyperCytautosampler is currently connected to a FACSCalibur, which allows for 3 color analysis.

HTS flow cytometry ideally suited for:
  • Large sample sets from patient studies
  • Hybridoma identification
  • Drug screening
Location: BBRB 557
 
srv-facsaria The FACSAria is a state of the art cell 4-laser sorter; 4 way sorting of up to 14 parameters is possible. The two machines have different filter and lasers set-ups.
Location: BBRB 557 and SHEL 271
srv-bioplex
The BioPlex suspension array system offers protein and nucleic acid researchers a reliable multiplex assay solution that permits analysis of up to 30biomolecules in a single sample.
BBRB 557
srv-bdlsr2 The LSR II is a state of the art 4-laser flow cytometer. Analysis can be up to 18 parameters; quantum-dot enabled.
Location: BBRB 557 and SHEL 271*
* An HTS system is available in SHEL 271
 
srv-facscalibur

FACSCalibur is a 2-laser flow cytometer that can detect up to four colors such as FITC, PE, PE-Cy5, or APC. A carousel system is available for automatic sample collection.
Location: SHEL 271
srv-facscan

FACScan is a user friendly flow cytometer that can detect up to three colors such, as FITC, PE, and PerCP or PE-Cy5.
Location: SHEL 271
srv-automacs The autoMACS™ Separator is a bench top instrument for high-speed automated cell sorting (Miltenyi) using magnetic beads. Positive and negative selection possible.
Location: SHEL 271


Fees


All Assisted Services Fees: $75 / hour

Self-Service Equipment Costs:
LSR II: $50 / hour
FACScan: $35 / hour
FACSCalibur: $35 / hour
BioPlex analysis: $50 / analyzed plate
HTS fees are negotiated on a per-project basis

Training Fees: Not all training has associated costs. Costs depend on the proficiency of the user and the type of equipment.

Contact the facility directly for more information or to schedule training for specific equipment.

Flow Cytometry Web Scheduling

Publications




1. HIV-1 latency and virus production from unintegrated genomes following direct infection of resting CD4 T cells. Chan CN, Trinité B, Lee CS, Mahajan S, Anand A, Wodarz D, Sabbaj S, Bansal A, Goepfert PA, Levy DN. Retrovirology. 2016 Jan 5;13(1) PMID number: 26728316 PMCID number: PMC4700562

2. Enhanced allogeneic cellular responses to mismatched HLA-B antigens results in more efficient killing of HIV infected cells. Sabbaj S, Scanlon N, Du VY, Wang Y, Tang J, Hunter E, Goepfert PA. J Acquir Immune Defic Syndr. 2015 Nov 19. PMID number: 26588705 PMCID number: not provided

3. Normal T-cell activation in elite controllers with preserved CD4+ T-cell counts. Bansal A, Sterrett S, Erdmann N, Westfall AO, Dionne-Odom J, Overton ET, Goepfert PA. AIDS. 2015 Nov;29(17):2245-54. PMID number: 26544698 PMCID number: not provided

4. Vaccination With Heterologous HIV-1 Envelope Sequences and Heterologous Adenovirus Vectors Increases T-Cell Responses to Conserved Regions: HVTN 083. Walsh SR, Moodie Z, Fiore-Gartland AJ, Morgan C, Wilck MB, Hammer SM, Buchbinder SP, Kalams SA, Goepfert PA, Mulligan MJ, Keefer MC, Baden LR, Swann EM, Grant S, Ahmed H, Li F, Hertz T, Self SG, Friedrich D, Frahm N, Liao HX, Montefiori DC, Tomaras GD, McElrath MJ, Hural J, Graham BS, Jin X; HVTN 083 Study Group and the NIAID HVTN. J Infect Dis. 2016 Feb 15;213(4):541-50.  PMID number: 26475930 PMCID number: PMC4721914

5. Enhanced Recognition of HIV-1 Cryptic Epitopes Restricted by HLA Class I Alleles Associated With a Favorable Clinical Outcome. Bansal A, Mann T, Sterrett S, Peng BJ, Bet A, Carlson JM, Goepfert PA. J Acquir Immune Defic Syndr. 2015 Sep 1;70(1):1-8. PMID number: 26322665 PMCID number: PMC4556167

6. HLA Class-II Associated HIV Polymorphisms Predict Escape from CD4+ T Cell Responses. Erdmann N, Du VY, Carlson J, Schaefer M, Jureka A, Sterrett S, Yue L, Dilernia D, Lakhi S, Tang J, Sidney J, Gilmour J, Allen S, Hunter E, Heath S, Bansal A, Goepfert PA. PLoS Pathog. 2015 Aug 24;11(8):e1005111. PMID number:26302050 PMCID number: PMC4547780

7. Stable Phenotypic Changes of the Host T Cells Are Essential to the Long-Term Stability of Latent HIV-1 Infection. Seu L, Sabbaj S, Duverger A, Wagner F, Anderson JC, Davies E, Wolschendorf F, Willey CD, Saag MS, Goepfert P, Kutsch O. J Virol. 2015 Jul;89(13):6656-72. PMID number: 25878110 PMCID number: PMC4468477

8. A Context-Dependent Role for IL-21 in Modulating the Differentiation, Distribution, and Abundance of Effector and Memory CD8 T Cell Subsets. Tian Y, Cox MA, Kahan SM, Ingram JT, Bakshi RK, Zajac AJ. J Immunol. 2016 Mar 1;196(5):2153-66. PMID number: 26826252 PMCID number: PMC4761492

9. A Critical Role of IL-21-Induced BATF in Sustaining CD8-T-Cell-Mediated Chronic Viral Control. Xin G, Schauder DM, Lainez B, Weinstein JS, Dai Z, Chen Y, Esplugues E, Wen R, Wang D, Parish IA, Zajac AJ, Craft J, Cui W. Cell Rep. 2015 Nov 10;13(6):1118-24. PMID number: 26527008 PMCID number: not provided

10. Colonization potential to reconstitute a microbe community in patients detected early after fecal microbe transplant for recurrent C. difficile. Kumar R, Maynard CL, Eipers P, Goldsmith KT, Ptacek T, Grubbs JA, Dixon P, Howard D, Crossman DK, Crowley MR, Benjamin WH Jr, Lefkowitz EJ, Weaver CT, Rodriguez JM, Morrow CD. BMC Microbiol. 2016 Jan 13;16(1):5. PMID number:26758906 PMCID number: PMC4711103

11. IRF4-Dependent and IRF4-Independent Pathways Contribute to DC Dysfunction in Lupus. Manni M, Gupta S, Nixon BG, Weaver CT, Jessberger R, Pernis AB. PLoS One. 2015 Nov 6;10(11):e0141927. PMID number: 26544714 PMCID number: PMC4636285

12. Skin-resident memory CD4+ T cells enhance protection against Leishmania major infection. Glennie ND, Yeramilli VA, Beiting DP, Volk SW, Weaver CT, Scott P. J Exp Med. 2015 Aug 24;212(9):1405-14. PMID number: 26216123 PMCID number: PMC4548053

13. Th17 cells give rise to Th1 cells that are required for the pathogenesis of colitis. Harbour SN, Maynard CL, Zindl CL, Schoeb TR, Weaver CT. Proc Natl Acad Sci U S A. 2015 Jun 2;112(22):7061-6. PMID number: 26038559 PMCID number: PMC4460486

14. Targeting CXCR4/SDF-1 axis by lipopolymer complexes of siRNA in acute myeloid leukemia. Landry B, Gül-Uludağ H, Plianwong S, Kucharski C, Zak Z, Parmar MB, Kutsch O, Jiang H, Brandwein J, Uludağ H. J Control Release. 2016 Feb 28;224:8-21. PMID number:26742943 PMCID number: not provided

15. RNAP II processivity is a limiting step for HIV-1 transcription independent of orientation to and activity of endogenous neighboring promoters. Kaczmarek Michaels K, Wolschendorf F, Schiralli Lester GM, Natarajan M, Kutsch O, Henderson AJ. Virology. 2015 Dec;486:7-14. PMID number:26379089 PMCID number: PMC4679410

16. The host cell side of latent HIV-1 infection. Seu L, Kutsch O. Oncotarget. 2015 Aug 21;6(24):19920-1.
 PMID number: 26343367 PMCID number: PMC4652956

17. Disulfiram and Copper Ions Kill Mycobacterium tuberculosis in a Synergistic Manner. Dalecki AG, Haeili M, Shah S, Speer A, Niederweis M, Kutsch O, Wolschendorf F. Antimicrob Agents Chemother. 2015 Aug;59(8):4835-44. PMID number:26033731 PMCID number: PMC4505271

18. T Follicular Helper Cell Plasticity Shapes Pathogenic T Helper 2 Cell-Mediated Immunity to Inhaled House Dust Mite. Ballesteros-Tato A, Randall TD, Lund FE, Spolski R, Leonard WJ, León B. Immunity. 2016 Feb 16;44(2):259-73. PMID number:26825674 PMCID number: PMC4758890

19. B Lymphocytes Are Required during the Early Priming of CD4+ T Cells for Clearance of Pneumocystis Infection in Mice. Opata MM, Hollifield ML, Lund FE, Randall TD, Dunn R, Garvy BA, Feola DJ. J Immunol. 2015 Jul 15;195(2):611-20. PMID number:26041535 PMCID number: PMC4491042

20. IL-7 signalling represses Bcl-6 and the TFH gene program. McDonald PW, Read KA, Baker CE, Anderson AE, Powell MD, Ballesteros-Tato A, Oestreich KJ. Nat Commun. 2016 Jan 8;7:10285. PMID number: 26743592 PMCID number: not provided

21. Interferon-induced mechanosensing defects impede apoptotic cell clearance in lupus. Li H, Fu YX, Wu Q, Zhou Y, Crossman DK, Yang P, Li J, Luo B, Morel LM, Kabarowski JH, Yagita H, Ware CF, Hsu HC, Mountz JD. J Clin Invest. 2015 Jul 1;125(7):2877-90. PMID number:26098211 PMCID number: PMC4563689

22. A Heterozygous RAB27A Mutation Associated with Delayed Cytolytic Granule Polarization and Hemophagocytic Lymphohistiocytosis. Zhang M, Bracaglia C, Prencipe G, Bemrich-Stolz CJ, Beukelman T, Dimmitt RA, Chatham WW, Zhang K, Li H, Walter MR, De Benedetti F, Grom AA, Cron RQ. J Immunol. 2016 Feb 15. pii: 1501284. PMID number:26880764 PMCID number: not provided

23. Inhibition of System Xc(-) Transporter Attenuates Autoimmune Inflammatory Demyelination. Evonuk KS, Baker BJ, Doyle RE, Moseley CE, Sestero CM, Johnston BP, De Sarno P, Tang A, Gembitsky I, Hewett SJ, Weaver CT, Raman C, DeSilva TM. J Immunol. 2015 Jul 15;195(2):450-63. PMCID number: PMC4490999

24. Pore-Forming Toxins Induce Macrophage Necroptosis during Acute Bacterial Pneumonia. González-Juarbe N, Gilley RP, Hinojosa CA, Bradley KM, Kamei A, Gao G, Dube PH, Bergman MA, Orihuela CJ. PLoS Pathog. 2015 Dec 11;11(12):e1005337. PMID number:26659062 PMCID number: PMC4676650

25. Chronic mTOR inhibition in mice with rapamycin alters T, B, myeloid, and innate lymphoid cells and gut flora and prolongs life of immune-deficient mice. Hurez V, Dao V, Liu A, Pandeswara S, Gelfond J, Sun L, Bergman M, Orihuela CJ, Galvan V, Padrón Á, Drerup J, Liu Y, Hasty P, Sharp ZD, Curiel TJ. Aging Cell. 2015 Dec;14(6):945-56. PMCID number: PMC4693453

26. Loss of NOX-Derived Superoxide Exacerbates Diabetogenic CD4 T-Cell Effector Responses in Type 1 Diabetes. Padgett LE, Anderson B, Liu C, Ganini D, Mason RP, Piganelli JD, Mathews CE, Tse HM. Diabetes. 2015 Dec;64(12):4171-83. PMID number:26269022 PMCID number: PMC4657579

27. Optimized Replicating Renilla Luciferase Reporter HIV-1 Utilizing Novel Internal Ribosome Entry Site Elements for Native Nef Expression and Function. Alberti MO, Jones JJ, Miglietta R, Ding H, Bakshi RK, Edmonds TG, Kappes JC, Ochsenbauer C. AIDS Res Hum Retroviruses. 2015 Dec;31(12):1278-96. PMID number:26101895 PMCID number: PMC4663642

28. Resolution of Chlamydia trachomatis Infection Is Associated with a Distinct T Cell Response Profile. Picard MD, Bodmer JL, Gierahn TM, Lee A, Price J, Cohane K, Clemens V, DeVault VL, Gurok G, Kohberger R, Higgins DE, Siber GR, Flechtner JB, Geisler WM. Clin Vaccine Immunol. 2015 Nov;22(11):1206-18 PMID number:26446421







Useful Links


A 12 minute video introduction to flow cytometry: A must see for beginners :
http://probes.invitrogen.com/resources/education/tutorials/4Intro_Flow/player.html 

Flow Cytometry – A Basic Introduction by Michael G. Ormerod
http://flowbook.denovosoftware.com/ 

Practical Flow Cytometry --- Howard Shapiro
http://www.beckmancoulter.com/wsrportal/wsr/research-and-discovery/products-and-services/flow-cytometry/practical-flow-cytometry/index.htm 

A useful Java applet put together by Joe Trotter of BD Biosciences. You can view the excitation and emission spectra of many common fluorescent dyes for help in panel design.
http://www.bdbiosciences.com/research/multicolor/tools/index.jsp 

An applet similar to the above that has all of the Invtrogen reagents, as well as common fluorchromes
http://www.invitrogen.com/site/us/en/home/support/Research-Tools/Fluorescence-SpectraViewer.html

For FlowJo users: http://flowjo.typepad.com/ 

Excellent information on flow cytometry applications, especially intracellular staining
http://maeckerlab.typepad.com/maeckerlab_weblog/maeckerlab_protocols_and_tools/ 

Invitrogen: The current home of the legendary Molecular Probes materials and lots of other useful information:
http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cell-and-Tissue-Analysis/Flow-Cytometry.html 

Cell Signaling
http://www.cellsignal.com/technologies/flow.html 

Millipore GUAVA
http://www.millipore.com/flowcytometry/fc/fchome&open&cid=U0021 

Intellicyt --- HyperCyt autosampler for high throughput flow cytometry (available in CFAR Core)
http://www.intellicyt.com/instruments/hypercyt-sampler

Contact Information


BBRB Location:

Director: Olaf Kutsch, PhD
Manager: Marion Spell, BS
E-mail: mlspell@uab.edu
Phone: (205) 975-3227

Bevill Basic Research Building
Room 557
845 19th Street South
Birmingham, Alabama, 35294

Shelby Location

Director: John D. Mountz, MD, PhD
Manager: Enid F. Keyser, BS
E-mail: efk@uab.edu
(205) 934-1362

Shelby Biomedical Research Building
Room 271
1825 University Blvd
Birmingham, Alabama, 35294