Analytical Cytometry

To advance our understanding of cancer etiology and to develop improved therapeutic approaches, it is critical for researchers to have the ability to routinely analyze and isolate discrete cell types from complex heterogeneous populations. The Analytical Cytometry Shared Resource (AC-SR) fulfills this vital function for COHCCC Members by providing services to define and isolate discrete immune system cells and cancer cells based on extracellular and intracellular protein expression, to quantify and isolate cells that express fluorescent markers, and to examine discrete cell growth states.

The Analytical Cytometry Shared Resource (AC-SR) provides flow cytometry analysis and sorting instrumentation (18 analyzers, seven sorters) to support the research goals of COHCCC. The AC-SR maintains analysis instruments (7-BD LSRII/Fortessa, Cytek Aurora, BD-Symphony, 3-ThermoFisher Attune NxT, 2-ACEA Quanteons, 1-ACEA Novocyte, 2-BD C6, and BC CyAn) that enable varying degrees of parameter detection for all COH sites. A mass cytometer, Fluidigm CyTOF-Helios, and a small particle analyzer (e.g., exosome and nanoparticle; NanoSight NS300) is also available. Notably, the Aurora, Symphony, and CyTOF instruments are new since the last funding cycle. These instruments allow for high parameter experiments, enabling highly detailed analysis and interrogation of cancer cells, cancer microenvironments, and immunological responses. All analysis instruments can be operated either by researchers or by SR staff. Six cell sorting instruments (BD Aria sorters) are located across COH sites of which 4 are available for self-operation by the trained end user. COHCCC has recently purchased an additional closed system cell sorter (Miltenyi Tyto) to assist with sorting of CAR-T cell products and oncolytic viral research needs. The SR is directed by Dr. Jeremy Stark, a Professor in the Department of Cancer Genetics and Epigenetics and supported by highly qualified staff that maintain and operate the equipment while also providing training to COHCCC researchers who wish to operate AC-SR instruments independently.

Other CCSG Data

Specific Aims for AC-SR:

Aim 1. Support flow cytometry experiments for the analysis of cells and small particles.

Aim 2. Maintain instrumentation for cell sorting and perform multiparameter sorting.

Aim 3. Provide training support for researchers to perform independent flow cytometry analysis and sorting within the AC-SR.

Members Utilization by % Revenue 2017–21: 93.6 Total (20 MCBC, 4.1 DCT, 24 CI, 45 HM, 0.26 CCPS)
Publications by Members: 172, 95 with Impact Factor >10
Grants Supported: 169Total (4 ACS, 12 CIRM, 8 DoD, 6 LLS, 18 NCI of 32NIH (1P01, 88R01, 5U01))

Using the Analytical Cytometry Facility

Resources for Grants

The Analytical Cytometry Core (ACC) comprises seven cell sorters, 16 analytical cytometers, CyTOF-Helios mass cytometer, and a nanoparticle analysis instrument.

Our cell sorters

ARIA III (Becton Dickinson) with four lasers (405nm, 488nm, 561nm and 633nm) and up to 15 fluorescent parameters. This sorter is contained in a biosafety cabinet.
ARIA II SORP (Becton Dickinson) with six lasers (355nm, 405nm, 457nm, 488nm, 561nm and 640nm) and up to 18 simultaneous fluorescent parameters. This sorter is contained in a biosafety cabinet.
ARIA Fusion (Becton Dickinson) with five lasers (355nm, 405nm, 488nm, 561nm and 640nm) and up to 18 simultaneous fluorescent parameters. These sorters are contained in a biosafety cabinet.
Tyto Cell Sorter with 3 lasers (405nm, 488nm, and 638nm) and 8 fluorescent detectors. Requires specific cartridges for operation.

Our analytical cytometers

Novocyte3005 with 3 laser (405nm, 488nm, 640nm) system equipped with 14 fluorescent detectors.
Quanteon 4025 with 4 laser system (405nm, 488nm, 561nm, 637nm) equipped with 25 fluorescent detectors.
Aurora with 5 laser system (355nm, 405nm, 488nm, 561nm, 640nm) full spectrum analyzer equipped with 64 fluorescent detectors.
Fortessa SORP (Becton Dickinson) with five lasers (355nm, 405nm, 488nm, 561nm and 640nm) and up to 18 simultaneous fluorescent parameters. Also available is an HTS module for 96-well sample delivery allowing for automation.
Fortessa X-20 (Becton Dickinson) with five lasers (355nm, 405nm, 488nm, 561nm and 640nm) and up to 18 simultaneous fluorescent parameters. Also available is an HTS module for 96-well sample delivery allowing for automation.
LSRII (Becton Dickinson) with five lasers (355nm, 405nm, 488nm, 561nm and 640nm) and up to 15 simultaneous fluorescent parameters.
FACSymphony (Becton Dickinson) with five high-powered lasers (355nm, 405nm, 488nm, 561nm and 640nm) and up to 28 simultaneous fluorescent parameters.
C6 (Becton Dickinson) with four fluorescent detectors and two lasers (488nm-3 detectors and 640nm-1 detector).
Attune cytometer (ThermoFisher) with four lasers (405nm, 488nm, 561nm and 640nm) and 14 simultaneous fluorescent parameters. It has sample delivery from tube or 96-well format integrated in the system (controlled by software).

Other instrumentation

NanoSight NS300 (Malvern) allows for the characterization of submicron size particles.
CyTOF-Helios (Fluidigm) utilizes metal-tagged antibodies or reagents to detect up to 50 simultaneous parameters on a per cell basis.

ACC also provides training for user operation of the instruments, basic flow theory, software, and data analysis for result generation and presentation. Other functions include application assistance, experimental design, and site license management for Flowjo and FCSexpress software.

Analytical Cytometry Services

The Analytical Cytometry Core uses flow cytometric principles to analyze and sort samples (cells or particles) based on fluorescence or physical properties.

Cell sorting
Flow cytometry analysis
Data analysis
Equipment training — analyzer
Equipment training — sorter
FCS Express and Flowjo software access
Software training
Technical assistance

Pricing and Scheduling

Current pricing can be found on iLab Solutions.

Meet the Team

Leadership

Jeremy Stark, Ph.D.

Core Director, Analytical Cytometry

Full bio

Lucy Brown, M.S.

Core Manager, Analytical Cytometry

Full bio

Core Staff

Alexander Spalla, B.S.
Senior Research Associate

Ni Feng, B.S.
Research Associate II

Shaun Hsueh, B.S.
Research Associate II

Sumana Majumdar, M.S.
Research Associate II

Contact the Team

To request or initiate services, please login or register via iLab Solutions. For any additional questions, please contact Lucy Brown.

[email protected]

iLab Solutions

Equipment

ARIA Fusion Cell Sorters

The Aria Fusion cell sorters (Becton Dickinson) are five-laser (355nm, 405nm, 488nm, 561nm and 640nm) with 18 simultaneous fluorescent detectors.

ARIA II SORP (Special Order Research Product)

The Aria II SORP (Becton Dickinson) is a six-laser system (355nm, 405nm, 457nm, 488nm, 561nm and 640nm) with a maximum of 17 simultaneous fluorescent detectors.

ARIA III Cell Sorter

The Aria III cell sorter (Becton Dickinson) is a four-laser system (405nm, 488nm, 561nm and 633nm) with a maximum of 15 fluorescent detectors.

ATTUNE NxT Analytical Cytometer

The Attune NxT (ThermoFisher) is a four-laser (405nm, 488nm, 561nm and 640nm) 14 fluorescent detector instrument.

Aurora

The Aurora is a 5 laser (355nm, 405nm, 488nm, 561nm, 640nm) full spectrum analyzer equipped with 64 fluorescent detectors. Full spectrum cytometers utilize the entire emission spectra of fluorophores which allows for greater data resolution and the utilization of more fluorophores simultaneously when compared with traditional flow cytometers. This system has the option of utilizing either 12x75mm 5mL FACS tubes or from 96-well plate formats f or data acquisition. This instrument is available for trained users to operate, or user samples can be analyzed by the core staff at an additional cost.

C6 Analytical Cytometer

The C6 (Becton Dickinson) is a two-laser (488nm and 640nm) system. Up to three fluorescent detectors are available for the 488nm laser. One detector is available for the 640nm laser. Trained users may operate this cytometer.

CyTOF-Helios Mass Cytometer

CyTOF-Helios (Fluidigm) utilizes mass cytometry, which uniquely combines time-of-flight mass spectrometry with metal-tagged antibodies or reagents to detect up to 50 simultaneous parameters on a per cell basis.

FACSymphony Analytical Cytometer

This Becton Dickinson cytometer has five lasers (355nm, 405nm, 488nm, 561nm and 640nm).

LSRII, Fortessa and FortessaX20 Analytical Cytometers

These instruments are five-laser analytical cytometers. The Becton Dickinson cytometers are special order research products equipped with 355nm (UV), 405nm, 488nm, 561nm and 640nm lasers.

Miltenyi Tyto Cell Sorter

The Miltenyi Tyto cell sorter is a microfluidics based system. It contains 3 lasers (405nm, 488nm, and 638nm) and 8 fluorescent detectors. Requires specific cartridges for operation.

Miltenyi Tyto Cell Sorter Optical Configuration

NanoSight NS300

NanoSight NS300 (Malvern) allows for the characterization of submicron-sized particles.

Novocyte 3005

The Novocyte is a 3 laser (405nm, 488nm, 640nm) system equipped with 14 fluorescent detectors. Capable of running samples from either 12x75mm 5mL tubes or from 96-well plate formats. This instruments is available for trained users to operate, or user samples can be analyzed by the core staff at an additional cost.

Novocyte 3005 optical configuration

Quanteon 4025

The Quanteon 4025 is a 4 laser system (405nm, 488nm, 561nm, 637nm) equipped with 25 fluorescent detectors. Capable of running samples from 12x75mm 5mL tubes, microfuge tubes, and from 96-well plate formats. These instruments are available for trained users to operate, or user samples can be analyzed by the core staff at an additional cost.

NC Quanteon optical configurations

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Our Publications

MiR-16 regulates crosstalk in NF-κB tolerogenic inflammatory signaling between myeloma cells and bone marrow macrophages. JCI insight, 4(21), e129348. (2019).

Khalife, J., Ghose, J., Martella, M., Viola, D., Rocci, A., Troadec, E., Terrazas, C., Satoskar, A. R., Gunes, E. G., Dona, A., Sanchez, J. F., Bergsagel, P. L., Chesi, M., Pozhitkov, A., Rosen, S., Marcucci, G., Keats, J. J., Hofmeister, C. C., Krishnan, A., Caserta, E., … Pichiorri, F.

Targeted Activation of Cystic Fibrosis Transmembrane Conductance Regulator. Molecular therapy : the journal of the American Society of Gene Therapy, 27(10), 1737–1748. (2019).

Villamizar, O., Waters, S. A., Scott, T., Saayman, S., Grepo, N., Urak, R., Davis, A., Jaffe, A., & Morris, K. V.

Extrafollicular CD4+ T-B interactions are sufficient for inducing autoimmune-like chronic graft-versus-host disease. Nature communications, 8(1), 978. (2017).

Deng, R., Hurtz, C., Song, Q., Yue, C., Xiao, G., Yu, H., Wu, X., Müschen, M., Forman, S., Martin, P. J., & Zeng, D.

C-NHEJ without indels is robust and requires synergistic function of distinct XLF domains. Nature communications, 9(1), 2484. (2018).

Bhargava, R., Sandhu, M., Muk, S., Lee, G., Vaidehi, N., & Stark, J. M.

PRMT1-mediated FLT3 arginine methylation promotes maintenance of FLT3-ITD+ acute myeloid leukemia. Blood, 134(6), 548–560. (2019).

He, X., Zhu, Y., Lin, Y. C., Li, M., Du, J., Dong, H., Sun, J., Zhu, L., Wang, H., Ding, Z., Zhang, L., Zhang, L., Zhao, D., Wang, Z., Wu, H., Zhang, H., Jiang, W., Xu, Y., Jin, J., Shen, Y., … Li, L.

Targeting cell membrane HDM2: A novel therapeutic approach for acute myeloid leukemia. Leukemia, 34(1), 75–86. (2020).

Wang, H., Zhao, D., Nguyen, L. X., Wu, H., Li, L., Dong, D., Troadec, E., Zhu, Y., Hoang, D. H., Stein, A. S., Al Malki, M., Aldoss, I., Lin, A., Ghoda, L. Y., McDonald, T., Pichiorri, F., Carlesso, N., Kuo, Y. H., Zhang, B., Jin, J., … Marcucci, G.

Dual Mechanisms of Action of Self-Delivering, Anti-HIV-1 FANA Oligonucleotides as a Potential New Approach to HIV Therapy. Molecular therapy. Nucleic acids, 17, 615–625. (2019).

Takahashi, M., Li, H., Zhou, J., Chomchan, P., Aishwarya, V., Damha, M. J., & Rossi, J. J.

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City of Hope is focused on basic and clinical research in cancer, diabetes and other life-threatening diseases.

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