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Flow and Immune Analysis Shared Resource

Contact Info:

Orla Maguire

Elm and Carlton Streets

Buffalo, NY 14263

https://www.roswellpark.org/shared-resources/flow-immune-analysis

Grants and Identifiers:

RRID: RRID:SCR_022313

Other Citation Identifiers

2P30CA016056-32

5R50CA211108-07

1S10OD018048-01

1S10OD025183-01

Instrumentation:

BD FACSAria II Cell Sorter

Cell sorter features hardware and software enhancements that improve overall ease of use, flexibility,and aseptic capability. Offers new options in lasers and nozzles to support more advanced multicolor applications. Built on fixed alignment technology. FACSAria II cell sorter is first generation of BD FACSAria system where flow cell is in true fixed alignment with laser, to reduce startup time and improve reproducibility. [Product Link]

RRID:SCR_018934

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BD LSRFortessa Fortessa Flow Cytometer

BD LSRFortessa cell analyzer offers the ultimate in choice for flow cytometry, providing power, performance, and consistency. Designed to be and expandable, the BD LSRFortessa has the flexibility to support the expanding needs of multicolor flow cytometry assays. The BD LSRFortessa system can be configured with up to 7 lasers*,blue, red, violet, UV and yellow-green. The instrument can accommodate the detection of up to 18 colors simultaneously with a defined set of optical filters that meet or exceed the majority of today's assay requirements. BD FACSDiva software controls the efficient setup, acquisition, and analysis of flow cytometry data from the BD LSRFortessa workstation. The software is common across BD FACS instrument families, including the BD FACSCanto cell analyzer and BD FACSAria cell sorter systems. [Product Link]

RRID:SCR_019601

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(Equipment No Longer Active At this Facility)

BD LSRFortessa X-20 Fortessa Flow Cytometer

BD LSRFortessa cell analyzer offers the ultimate in choice for flow cytometry, providing power, performance, and consistency. Designed to be and expandable, the BD LSRFortessa has the flexibility to support the expanding needs of multicolor flow cytometry assays. The BD LSRFortessa system can be configured with up to 7 lasers*,blue, red, violet, UV and yellow-green. The instrument can accommodate the detection of up to 18 colors simultaneously with a defined set of optical filters that meet or exceed the majority of today's assay requirements. BD FACSDiva software controls the efficient setup, acquisition, and analysis of flow cytometry data from the BD LSRFortessa workstation. The software is common across BD FACS instrument families, including the BD FACSCanto cell analyzer and BD FACSAria cell sorter systems. [Product Link]

RRID:SCR_019600

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Cytek Aurora Spectral Analyzer

Prodigy incorporating a combination of patent-pending technologies that takes flow cytometry to the next level of performance and flexibility. With up to five lasers, three scattering channels, and 64 fluorescence channels, the Aurora suits every laboratory's needs, from to high complexity applications. A paradigm shifting optical design provides unprecedented flexibility, enabling the use of a wide array of new fluorochrome combinations without reconfiguring your system for each application. The state-of-the-art optics and low-noise electronics provide excellent sensitivity and resolution. Flat-top beam profiles, combined with a ly designed fluidics system, translate to outstanding performance at high sample flow rates. [Product Link]

RRID:SCR_019826

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ImageStream-X, Image Analyzer (4 lasers/10 colors) Multiple

ImageStreamX Mk II System is a benchtop, multispectral, imaging flow cytometer designed for the acquisition of up to 12 channels of cellular imagery. By collecting large numbers of digital images per sample and providing a numerical representation of image-based features, the ImageStreamX Mk II combines the per cell information content provided by standard microscopy with the statistical significance afforded by large sample sizes common to standard flow cytometry. With the ImageStreamX Mk II System, fluorescence intensity measurements are acquired as with a conventional flow cytometer; however, the best applications for the ImageStreamX Mk II take advantage of the system's imaging abilities to locate and quantitate the distribution of signals on in or between cells. [Product Link]

RRID:SCR_020142

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Sony MA900 Multi-Application Cell Sorter

MA900 system include patented micro fluidic chip-based design, comprehensive fluidic controls, and advanced automation that simplifies operation to make sorting less subjective and improve reliability. Provides up to 14 parameters from 4 lasers with possibility to choose from 4 excitation lasers—488 nm, 405 nm, 561 nm, and 638 nm—on two beam spots. Free-form PMTs enable detection of fluorescence signals from each beam spot, allowing detection of up to 12 fluorescence parameters and two scatter parameters. [Product Link]

RRID:SCR_026300

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Akoya Phenocycler Fusion

Spatial biology solution that enables ultrahigh-plex spatial phenotyping of whole slides at single-cell resolution by integrating automated fluidics and iterative imaging.

Particle Metrix ZetaView

Small particle analyzer

Services Provided:

Akoya PhenoCycler Fusion

Analysis Workstations

Assay Development

Assays and Measurements

Cell Sorting

Confocal Microscopy

Consultation

Cytokine Assays

Data Analysis

ELISA

Exosomes Characterisation

Experimental Design

Flow Cytometric Analysis

Flow Cytometry

Flow Cytometry Data Analysis

Imaging Flow Cytomtry

Immune Monitoring

Luminex Bead Based Cytokine Assays

Luminex Services

Multi-color Flow Cytometry

Multiplexed Immunofluorescence

Phenotyping

Spectral Flow Cytometry

Staining

Tissue Culture

Relevant Publications:

1.) Dong B, Obermajer N, Tsuji T, Matsuzaki J, Bonura CM, Sander C, Withers H, Long MD, Chavel C, Olejniczak SH, Minderman H, Kirkwood JM, Edwards RP, Storkus WJ, Romero P, Kalinski P (2024 Oct 1). NK Receptor Signaling Lowers TCR Activation Threshold, Enhancing Selective Recognition of Cancer Cells by TAA-Specific CTLs. Cancer immunology research, 12(10), 1421-1437. . ID: 38949179.


2.) Alaklabi S, Maguire O, Pattnaik H, Zhang Y, Chow J, Wang J, Minderman H, Iyer R (2024 Sep 7). Immune Cell Molecular Pharmacodynamics of Lanreotide in Relation to Treatment Response in Patients with Gastroenteropancreatic Neuroendocrine Tumors. Cancers, 16(17), . . ID: 39272962.


3.) Maguire OR, Smokers IBA, Oosterom BG, Zheliezniak A, Huck WTS (2024 Mar 20). A Prebiotic Precursor to Life Journal of the American Chemical Society, 146(11), 7839-7849. . ID: 38448161.


4.) McCarthy PL, Attwood KM, Liu X, Chen GL, Minderman H, Alousi A, Bashey A, Lowsky R, Miklos DB, Hansen J, Westervelt P, Yanik G, Waller EK, Howard A, Blazar BR, Wallace PK, Reshef R, Horowitz MM, Maziarz RT, Levine JE, Mohammadpour H (2024 Mar). Galectin-3 predicts acute GvHD and overall mortality post reduced intensity allo-HCT: a BMT-CTN biorepository study. Bone marrow transplantation, 59(3), 334-343. . ID: 38110620.


5.) Seon BK, Okazaki M, Duzen J, Matsuno F, Goey AKL, Maguire O (2024 Jan). Identification of unique molecular heterogeneity of human CD79, the signaling component of the human B cell antigen receptor (BCR), and synergistic potentiation of the CD79-targeted therapy of B cell tumors by co-targeting of CD79a and CD79b. Leukemia research, 136(), 107436. . ID: 38232613.


6.) Gandhi S, Opyrchal M, Grimm MJ, Slomba RT, Kokolus KM, Witkiewicz A, Attwood K, Groman A, Williams L, Tarquini ML, Wallace PK, Soh KT, Minderman H, Maguire O, O (2023 Nov). Systemic infusion of TLR3-ligand and IFN-α in patients with breast cancer reprograms local tumor microenvironments for selective CTL influx. Journal for immunotherapy of cancer, 11(11), . . ID: 37963636.


7.) Dong B, Obermajer N, Tsuji T, Matsuzaki J, Bonura C, Withers H, Long M, Chavel C, Olejniczak SH, Minderman H, Edwards RP, Storkus WJ, Romero P, Kalinski P (2023 Oct 19). NK Receptors Replace CD28 As the Dominant Source of Signal 2 for Cognate Recognition of Cancer Cells by TAA-specific Effector CD8(+) T Cells. Research square, (), . . ID: 37886562.


8.) Tornatore KM, Attwood K, Brazeau D, Sprowl J, Chang S, Gundroo A, Minderman H, Venuto RC (2023 Feb). Comparison of P-glycoprotein function in peripheral blood mononuclear cells ex vivo in stable Black and White male and female kidney transplant recipients. Clinical and translational science, 16(2), 184-192. . ID: 36352830.


9.) Elshoury A, Maguire O, Conway A, Tario J, Soh KT, Ross M, Hahn T, Becker J, Wallace P, McCarthy PL, Minderman H, Chen GL (2023 Feb). Short-Term Storage of Mobilized Peripheral Blood Stem Cells in a Closed System Changes the Microenvironment and May Affect the Quantity of CD34(+) and CD34(+)CD38(-)CD45RA(-)CD90(+) Cells. Transplantation and cellular therapy, 29(2), 112.e1-112.e9. . ID: 36436782.


10.) Babiker H, Borazanci E, Subbiah V, Agarwala S, Algazi A, Schachter J, Lotem M, Maurice-Dror C, Hendler D, Rahimian S, Minderman H, Haymaker C, Mahadevan D, Bernatchez C, Murthy R, Hultsch R, Kaplan N, Woodhead G, Hennemeyer C, Chunduru S, Anderson PM, Diab A, Puzanov I (2022 Dec 1). Tilsotolimod Exploits the TLR9 Pathway to Promote Antigen Presentation and Type 1 IFN Signaling in Solid Tumors: A Multicenter International Phase I/II Trial (ILLUMINATE-101). Clinical cancer research : an official journal of the American Association for Cancer Research, 28(23), 5079-5087. . ID: 35917516.


11.) Rosenberg CA, Bill M, Maguire O, Petersen MA, Kjeldsen E, Hokland P, Ludvigsen M (2022 Mar). Imaging flow cytometry reveals a subset of TdT negative T-ALL blasts with very low forward scatter on conventional flow cytometry. Cytometry. Part B, Clinical cytometry, 102(2), 107-114. doi: 10.1002/cyto.b.22035. ID: 34648681.


12.) Mistarz A, Graczyk M, Winkler M, Singh PK, Cortes E, Miliotto A, Liu S, Long M, Yan L, Stablewski A, O (2021 Dec 17). Induction of cell death in ovarian cancer cells by doxorubicin and oncolytic vaccinia virus is associated with CREB3L1 activation. Molecular therapy oncolytics, 23(), 38-50. . ID: 34632049.


13.) Fountzilas C, Bajor DL, Mukherjee S, Saltzman J, Witkiewicz AK, Maguire O, Minderman H, Nambiar R, Rosenheck HR, Knudsen ES, Muhitch JB, Abrams SI, Wang C, Hutson AD, Attwood K, Hicks KA, Jurcevic JA, Kalinski P, Iyer R, Boland PM (2021 Dec 15). Phase Ib/II Study of Cetuximab plus Pembrolizumab in Patients with Advanced RAS Wild-Type Colorectal Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(24), 6726-6736. . ID: 34645646.


14.) Bhatta M, Shenoy GN, Loyall JL, Gray BD, Bapardekar M, Conway A, Minderman H, Kelleher RJ Jr, Carreno BM, Linette G, Shultz LD, Odunsi K, Balu-Iyer SV, Pak KY, Bankert RB (2021 Oct). Novel phosphatidylserine-binding molecule enhances antitumor T-cell responses by targeting immunosuppressive exosomes in human tumor microenvironments. Journal for immunotherapy of cancer, 9(10), . . ID: 34599030.


15.) Ngai LL, Ma CY, Maguire O, Do AD, Robert A, Logan AC, Griffiths EA, Nemeth MJ, Green C, Pourmohamad T, van Kuijk BJ, Snel AN, Kwidama ZW, Venniker-Punt B, Cooper J, Manz MG, Gjertsen BT, Smit L, Ossenkoppele GJ, Janssen JJWM, Cloos J, Sumiyoshi T (2021 Sep). Bimodal expression of potential drug target CLL-1 (CLEC12A) on CD34+ blasts of AML patients. European journal of haematology, 107(3), 343-353. . ID: 34053123.


16.) Oba T, Makino K, Kajihara R, Yokoi T, Araki R, Abe M, Minderman H, Chang AE, Odunsi K, Ito F (2021 May). In situ delivery of iPSC-derived dendritic cells with local radiotherapy generates systemic antitumor immunity and potentiates PD-L1 blockade in preclinical poorly immunogenic tumor models. Journal for immunotherapy of cancer, 9(5), . . ID: 34049930.


17.) Shu S, Allen CL, Benjamin-Davalos S, Koroleva M, MacFarland D, Minderman H, Ernstoff MS (2021). A Rapid Exosome Isolation Using Ultrafiltration and Size Exclusion Chromatography (REIUS) Method for Exosome Isolation from Melanoma Cell Lines. Methods in molecular biology (Clifton, N.J.), 2265(), 289-304. . ID: 33704723.


18.) Sribenja S, Maguire O, Attwood K, Buas MF, Palmer JR, Ambrosone CB, Higgins MJ (2021). Deletion of Foxa1 in the mouse mammary gland results in abnormal accumulation of luminal progenitor cells: a link between reproductive factors and ER-/TNBC breast cancer? American journal of cancer research, 11(6), 3263-3270. . ID: 34249460.


19.) Oba T, Long MD, Keler T, Marsh HC, Minderman H, Abrams SI, Liu S, Ito F (2020 Oct 27). Overcoming primary and acquired resistance to anti-PD-L1 therapy by induction and activation of tumor-residing cDC1s. Nature communications, 11(1), 5415. . ID: 33110069.


20.) Shu S, Matsuzaki J, Want MY, Conway A, Benjamin-Davalos S, Allen CL, Koroleva M, Battaglia S, Odunsi A, Minderman H, Ernstoff MS (2020 Oct). An Immunosuppressive Effect of Melanoma-derived Exosomes on NY-ESO-1 Antigen-specific Human CD8(+) T Cells is Dependent on IL-10 and Independent of BRAF(V600E) Mutation in Melanoma Cell Lines. Immunological investigations, 49(7), 744-757. . ID: 32799717.


21.) Iyer RV, Konda B, Fountzilas C, Mukherjee S, Owen D, Attwood K, Wang C, Maguire O, Minderman H, Suffren SA, Hicks K, Wilton J, Bies R, Casucci D, Reidy-Lagunes D, Shah M (2020 Aug 15). Multicenter phase 2 trial of nintedanib in advanced nonpancreatic neuroendocrine tumors. Cancer, 126(16), 3689-3697. . ID: 32525561.


22.) Utley A, Chavel C, Lightman S, Holling GA, Cooper J, Peng P, Liu W, Barwick BG, Gavile CM, Maguire O, Murray-Dupuis M, Rozanski C, Jordan MS, Kambayashi T, Olejniczak SH, Boise LH, Lee KP (2020 Jun 23). CD28 Regulates Metabolic Fitness for Long-Lived Plasma Cell Survival. Cell reports, 31(12), 107815. . ID: 32579940.


23.) Shu S, Yang Y, Allen CL, Hurley E, Tung KH, Minderman H, Wu Y, Ernstoff MS (2020). Purity and yield of melanoma exosomes are dependent on isolation method. Journal of extracellular vesicles, 9(1), 1692401. . ID: 31807236.


24.) Cossarizza A, Chang HD, Radbruch A, Acs A, Adam D, Adam-Klages S, Agace WW, Aghaeepour N, Akdis M, Allez M, Almeida LN, Alvisi G, Anderson G, Andrä I, Annunziato F, Anselmo A, Bacher P, Baldari CT, Bari S, Barnaba V, Barros-Martins J, Battistini L, Bauer W, Baumgart S, Baumgarth N, Baumjohann D, Baying B, Bebawy M, Becher B, Beisker W, Benes V, Beyaert R, Blanco A, Boardman DA, Bogdan C, Borger JG, Borsellino G, Boulais PE, Bradford JA, Brenner D, Brinkman RR, Brooks AES, Busch DH, Büscher M, Bu (2019 Oct). Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition). European journal of immunology, 49(10), 1457-1973. . ID: 31633216.


25.) Minderman H (2019 Sep). Simultaneous Analysis of Phenotype and Cytogenetics Using Imaging Flow Cytometry: Time to Teach Old Dogs New Tricks. Cytometry. Part A : the journal of the International Society for Analytical Cytology, 95(9), 943-945. . ID: 31006975.


26.) Iyer RV, Maguire O, Kim M, Curtin LI, Sexton S, Fisher DT, Schihl SA, Fetterly G, Menne S, Minderman H (2019 May 16). Dose-Dependent Sorafenib-Induced Immunosuppression Is Associated with Aberrant NFAT Activation and Expression of PD-1 in T Cells. Cancers, 11(5), . . ID: 31100868.


Description:

This state-of-the-art facility provides advanced flow cytometric and morphology services at cellular and subcellular levels of resolution. The services offered by the Flow and Immune Analysis Shared Resource (FIASR) include investigator access to equipment 24/7, education and consultation services, comprehensive sample processing, data acquisition and data analysis services, luminex cytokine, chemokine and growth factor quantification, and core flow cytometry services for investigator and biotech sponsored clinical trials.