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Core FC

Core H redThe UAB Flow Cytometry Facility is a component of the new CFAR Basic and Translational Sciences Core and 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.)       Moylan DC, Goepfert PA, Kempf MC, Saag MS, Richter HE, Mestecky J, Sabbaj S. Diminished CD103 (αEβ7) Expression on Resident T Cells from the Female Genital Tract of HIV-Positive Women. Pathog Immun. 2016 Fall-Winter;1(2):371-387. doi: 10.20411/pai.v1i2.166.
PMID: 28164171
PMCID: PMC5288734


2.)       Hel Z, Xu J, Denning WL, Helton ES, Huijbregts RP, Heath SL, Overton ET, Christmann BS, Elson CO, Goepfert PA, Mestecky J. Dysregulation of Systemic and Mucosal Humoral Responses to Microbial and Food Antigens as a Factor Contributing to Microbial Translocation and Chronic Inflammation in HIV-1 Infection. PLoS Pathog. 2017 Jan 26;13(1):e1006087. doi: 10.1371/journal.ppat.1006087.
PMID: 28125732
PMCID: PMC5268400


3.)       Prentice HA, Lu H, Price MA, Kamali A, Karita E, Lakhi S, Sanders EJ, Anzala O, Allen S, Goepfert PA, Hunter E, Gilmour J, Tang J. Dynamics and Correlates of CD8 T-Cell Counts in Africans with Primary Human Immunodeficiency Virus Type 1 Infection J Virol. 2016 Oct 28;90(22):10423-10430.
PMID: 27630231
PMCID: PMC5105675


4.)       Mónaco DC, Dilernia DA, Fiore-Gartland A, Yu T, Prince JL, Dennis KK, Qin K, Schaefer M, Claiborne DT, Kilembe W, Tang J, Price MA, Farmer P, Gilmour J, Bansal A, Allen S, Goepfert P, Hunter E. Balance between transmitted HLA preadapted and nonassociated polymorphisms is a major determinant of HIV-1 disease progression. J Exp Med. 2016 Sep 19;213(10):2049-63. doi: 10.1084/jem.20151984.
PMID: 27551154
PMCID: PMC5030801


5.)       Carlson JM, Du VY, Pfeifer N, Bansal A, Tan VY, Power K, Brumme CJ, Kreimer A, DeZiel CE, Fusi N, Schaefer M, Brockman MA, Gilmour J, Price MA, Kilembe W, Haubrich R, John M, Mallal S, Shapiro R, Frater J, Harrigan PR, Ndung'u T, Allen S, Heckerman D, Sidney J, Allen TM, Goulder PJ, Brumme ZL, Hunter E, Goepfert PA. Impact of pre-adapted HIV transmission. Nat Med. 2016 Jun;22(6):606-13. doi: 10.1038/nm.4100.
PMID: 27183217
PMCID: PMC4899163


6.)       Jordan SJ, Gupta K, Ogendi BM, Bakshi RK, Kapil R, Press CG, Sabbaj S, Lee JY, Geisler WM. The Predominant CD4+ Th1 Cytokine Elicited to Chlamydia trachomatis Infection in Women is Tumor Necrosis Factor-Alpha, not Interferon-Gamma. Clin Vaccine Immunol. 2017 Jan 18. pii: CVI.00010-17. doi: 10.1128/CVI.00010-17. [Epub ahead of print]
PMID: 28100498
PMCID: Not assigned


7.)       Matthews QL, Farrow AL, Rachakonda G, Gu L, Nde P, Krendelchtchikov A, Pratap S, Sakhare SS, Sabbaj S, Lima MF, Villalta F. Epitope Capsid-Incorporation: New Effective Approach for Vaccine Development for Chagas Disease. Pathog Immun. 2016 Fall-Winter;1(2):214-233.
PMID: 27709126
PMCID: PMC5046838


8.)       Schaaf K, Smith SR, Duverger A, Wagner F, Wolschendorf F, Westfall AO, Kutsch O, Sun J. Mycobacterium tuberculosis exploits the PPM1A signaling pathway to block host macrophage apoptosis. Sci Rep. 2017 Feb 8;7:42101. doi: 10.1038/srep42101.
PMID: 28176854
PMCID: PMC5296758


9.)       Schaaf K, Hayley V, Speer A, Wolschendorf F, Niederweis M, Kutsch O, Sun J. A Macrophage Infection Model to Predict Drug Efficacy Against Mycobacterium Tuberculosis. Assay Drug Dev Technol. 2016 Aug;14(6):345-54. doi: 10.1089/adt.2016.717.
PMID: 27327048
PMCID: PMC4991579


10.)    Yoder AC, Guo K, Dillon SM, Phang T, Lee EJ, Harper MS, Helm K, Kappes JC, Ochsenbauer C, McCarter MD, Wilson CC, Santiago ML. The transcriptome of HIV-1 infected intestinal CD4+ T cells exposed to enteric bacteria. PLoS Pathog. 2017 Feb 27;13(2):e1006226. doi: 10.1371/journal.ppat.1006226. [Epub ahead of print]
PMID: 28241075
PMCID: Not assigned


11.)    Rodriguez-Garcia M, Shen Z, Barr FD, Boesch AW, Ackerman ME, Kappes JC, Ochsenbauer C, Wira CR. Dendritic cells from the human female reproductive tract rapidly capture and respond to HIV. Mucosal Immunol. 2016 Aug 31. doi: 10.1038/mi.2016.72. [Epub ahead of print]
PMID: 27579858
PMCID: PMC5332537


12.)    Yang R, Masters AR, Fortner KA, Champagne DP, Yanguas-Casás N, Silberger DJ, Weaver CT, Haynes L, Rincon M. J IL-6 promotes the differentiation of a subset of naive CD8+ T cells into IL-21-producing B helper CD8+ T cells. Exp Med. 2016 Oct 17;213(11):2281-2291.
PMID: 27670591
PMCID: PMC5068236


13.)    Clement M, Marsden M, Stacey MA, Abdul-Karim J, Gimeno Brias S, Costa Bento D, Scurr MJ, Ghazal P, Weaver CT, Carlesso G, Clare S, Jones SA, Godkin A, Jones GW, Humphreys IR. Cytomegalovirus-Specific IL-10-Producing CD4+ T Cells Are Governed by Type-I IFN-Induced IL-27 and Promote Virus Persistence. PLoS Pathog. 2016 Dec 7;12(12):e1006050. doi: 10.1371/journal.ppat.1006050.
PMID: 27926930
PMCID: PMC5142785


14.)    Dalecki AG, Malalasekera AP, Schaaf K, Kutsch O, Bossmann SH, Wolschendorf F. Combinatorial phenotypic screen uncovers unrecognized family of extended thiourea inhibitors with copper-dependent anti-staphylococcal activity. Metallomics. 2016 Apr;8(4):412-21. doi: 10.1039/c6mt00003g.
PMID: 26935206
PMCID: 4838501


15.)    Sun J, Schaaf K, Duverger A, Wolschendorf F, Speer A, Wagner F, Niederweis M, Kutsch O. Protein phosphatase, Mg2+/Mn2+-dependent 1A controls the innate antiviral and antibacterial response of macrophages during HIV-1 and Mycobacterium tuberculosis infection. Oncotarget. 2016 Mar 29;7(13):15394-409. doi: 10.18632/oncotarget.8190.
PMID: 27004401
PMCID: PMC4941249


16.)    Shah S, Dalecki AG, Malalasekera AP, Crawford CL, Michalek SM, Kutsch O, Sun J, Bossmann SH, Wolschendorf F. 8-Hydroxyquinolines Are Boosting Agents of Copper-Related Toxicity in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2016 Sep 23;60(10):5765-76. doi: 10.1128/AAC.00325-16.
PMID: 27431227
PMCID: PMC5038248


17.)    Munguía-Fuentes R, Yam-Puc JC, Silva-Sánchez A, Marcial-Juárez E, Gallegos-Hernández IA, Calderón-Amador J, Randall TD, Flores-Romo L. Immunization of Newborn Mice Accelerates the Architectural Maturation of Lymph Nodes, But AID-Dependent IgG Responses Are Still Delayed Compared to the Adult. Front Immunol. 2017 Jan 19;8:13. doi: 10.3389/fimmu.2017.00013.
PMID: 28154564
PMCID: PMC5243854


18.)    Silva-Sanchez A, Randall TD. Fugue G Minor: Getting the Lymph Node Ensemble Together with Circadian Rhythm. Immunity. 2017 Jan 17;46(1):6-8. doi: 10.1016/j.immuni.2017.01.001.
PMID: 28099865
PMCID: Not assigned


19.)    Ballesteros-Tato A, Randall TD, Lund FE, Spolski R, Leonard WJ, León B. T Follicular Helper Cell Plasticity Shapes Pathogenic T Helper 2 Cell-Mediated Immunity to Inhaled House Dust Mite. Immunity. 2016 Feb 16;44(2):259-73. doi: 10.1016/j.immuni.2015.11.017.
PMID: 26825674
PMCID: PMC4758890
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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: 
E-mail: 
(205) 934-1362

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