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Immuno-Oncology Research Using Flow Cytometry
Immuno-oncology is the study of the immune system within a cancer environment [1]. Significant strides to reduce tumor burden have been made in this field with the development of antibodies, biologics, and adoptive T cell therapies. Current immuno-oncology needs include generating new therapeutics for better efficacy and safety through finding new targets and improving drug design.
Thermo Fisher Scientific has developed a wide variety of research tools to help you with your therapeutics research and development endeavors. Learn what flow cytometric reagents and analysis offer in a number of immuno-oncology research areas, including investigations into the tumor microenvironment, immune checkpoint analysis, and CAR T characterization.
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Download Immune guide
Learn more: Active areas of immuno-oncology research
Tumor microenvironments are composed of cancer, stromal, immune and other cell types [2]
Immune checkpoints are cell pathways crucial in maintaining a normal immune response and protecting tissues from damage when the immune system is activated [3]
CAR T cell engineering involves genetically modifying an individual’s own T cells to target antigens selectively expressed on cancer cells [4]
How many cells do you need?Try the rare events calculator
Flow cytometry: a useful tool in immuno-oncology research
Combining reagents and instruments can generate more complete and complex information about the immune system’s role in cancer. Learn about immuno-oncology workflows designed to enable you to spend less time learning protocols and more time performing experiments.
Prepare single-cell suspensions from tumor, blood, and cultured samples. Eliminate tedious isolation and expansion steps by using innovative and high-quality reagents.
Highlights include:
- State-of-the-art Dynabeads activation and expansion technology mimics in vivo T cell activation
- Fixation kits are compatible with analysis of most cellular antigens
- Proven protocols to prepare cells from both culture and in vivo samples
Thermo Fisher Scientific—including the eBioscience product portfolio—offers over 10,000 primary antibodies for flow cytometry use. The wide range of fluorochrome-conjugated antibodies identifies human, mouse, rat, or non-human primate cell antigens and can be multiplexed with our other assays.
Highlights include:
- Specifically developed, validated, and manufactured for flow cytometry applications
- Large number of antibodies conjugated to popular dyes including, FITC, and APC, Alexa Fluor, eFluor, and the new Super Bright polymer dyes
- Technical support and online tools to help build multicolor panels
Learn more at thermofisher.com/flowantibodies
The novel PrimeFlow assay methodology employs a proprietary fluorescent in situ hybridization (FISH) and branched DNA (bDNA) signal amplification technique that enables the detection of up to four RNA transcripts in a single cell and can be combined with standard flow cytometry antibody staining.
Highlights include:
- Superior analysis of gene expression as cells change over time or in response to stimuli
- High-throughput ready with optimized multi-well kits and probes for many RNA targets
- Assay for any protein when no antibody is available
Learn more at thermofisher.com/primeflow
Proliferation measurements are typically made based on DNA synthesis or on cellular metabolism parameters. Assays can report cell health, genotoxicity, or inhibition of tumor cell growth during drug development. CellTrace assays and Click-iT EdU offer sensitive reagents to measure proliferation in many cell types.
Highlights include:
- Quantitation of newly synthesized DNA
- Detection without denaturation of DNA
- Compatibility with sensitive R-PE tandems and fluorescent proteins
- Fast detection, in as little as 60 minutes
- An alternative to the cumbersome BrdU assay
Understanding the mechanisms of cell death and survival can represent a critical aspect of toxicological profiling and drug discovery. Invitrogen reagents and assays are designed to effectively study changes in the plasma membrane, the mitochondria, caspase activity, and DNA fragmentation and chromatin condensation as a result of apoptosis.
Highlights include:
- Many assays to detect different targets found in early-to-late events
- Range of fluorescent options to be excited from multiple laser excitation sources
- Assays are compatible and used in multiplexing experiments
Dead cells can give false-positive results as they nonspecifically bind to many reagents. Removing dead cells is a critical step for obtaining accurate flow cytometry results. Invitrogen cell viability reagents minimize the number steps to measure the percentage of live and dead cells.
Highlights include:
- A wide range of dyes spanning the spectrum and used in many fluorescence channels to fit into multicolor panels
- Fixable and non-fixable options
The Attune Flow Cytometers are high-performance instruments with features, capabilities and specifications that support the demands of researchers at the frontier of immuno-oncology research in the fight against cancer.
Highlights include:
- Faster acquisition of blood and tumor samples
- Less clogging with difficult and precious samples
- Intuitive software for easier compensation
Learn more at thermofisher.com/attune
Simultaneously detect multiple immune-oncology checkpoint proteins with panels designed to run on the Luminex platform. Quantification of soluble checkpoint molecules is a promising new approach that can also be combined with the assessment of cytokines and chemokines in multiplex immunoassays to support research in the field of immuno-oncology and cancer immunology.
Highlights include:
- Human Immuno-oncology Checkpoint panel (14-plex) for the Luminex platform
- Antibodies for key Immune checkpoint markers
- ELISA assays for Immune checkpoint markers
Efficiently use starting material by measuring up to 80 genes of interest in a single well in a 96- or 384-well plate. Based on the Luminex platform, QuantiGene Plex forgoes RNA purification and follows a streamlined ELISA-like workflow.
Highlights include:
- Direct-from-lysate measurement; no RNA purification required
- Works well with FFPE samples containing crosslinked, degraded RNA
- Multiplex up to 80 genes per well
Prepare single-cell suspensions from tumor, blood, and cultured samples. Eliminate tedious isolation and expansion steps by using innovative and high-quality reagents.
Highlights include:
- State-of-the-art Dynabeads activation and expansion technology mimics in vivo T cell activation
- Fixation kits are compatible with analysis of most cellular antigens
- Proven protocols to prepare cells from both culture and in vivo samples
Thermo Fisher Scientific—including the eBioscience product portfolio—offers over 10,000 primary antibodies for flow cytometry use. The wide range of fluorochrome-conjugated antibodies identifies human, mouse, rat, or non-human primate cell antigens and can be multiplexed with our other assays.
Highlights include:
- Specifically developed, validated, and manufactured for flow cytometry applications
- Large number of antibodies conjugated to popular dyes including, FITC, and APC, Alexa Fluor, eFluor, and the new Super Bright polymer dyes
- Technical support and online tools to help build multicolor panels
Learn more at thermofisher.com/flowantibodies
The novel PrimeFlow assay methodology employs a proprietary fluorescent in situ hybridization (FISH) and branched DNA (bDNA) signal amplification technique that enables the detection of up to four RNA transcripts in a single cell and can be combined with standard flow cytometry antibody staining.
Highlights include:
- Superior analysis of gene expression as cells change over time or in response to stimuli
- High-throughput ready with optimized multi-well kits and probes for many RNA targets
- Assay for any protein when no antibody is available
Learn more at thermofisher.com/primeflow
Proliferation measurements are typically made based on DNA synthesis or on cellular metabolism parameters. Assays can report cell health, genotoxicity, or inhibition of tumor cell growth during drug development. CellTrace assays and Click-iT EdU offer sensitive reagents to measure proliferation in many cell types.
Highlights include:
- Quantitation of newly synthesized DNA
- Detection without denaturation of DNA
- Compatibility with sensitive R-PE tandems and fluorescent proteins
- Fast detection, in as little as 60 minutes
- An alternative to the cumbersome BrdU assay
Understanding the mechanisms of cell death and survival can represent a critical aspect of toxicological profiling and drug discovery. Invitrogen reagents and assays are designed to effectively study changes in the plasma membrane, the mitochondria, caspase activity, and DNA fragmentation and chromatin condensation as a result of apoptosis.
Highlights include:
- Many assays to detect different targets found in early-to-late events
- Range of fluorescent options to be excited from multiple laser excitation sources
- Assays are compatible and used in multiplexing experiments
Dead cells can give false-positive results as they nonspecifically bind to many reagents. Removing dead cells is a critical step for obtaining accurate flow cytometry results. Invitrogen cell viability reagents minimize the number steps to measure the percentage of live and dead cells.
Highlights include:
- A wide range of dyes spanning the spectrum and used in many fluorescence channels to fit into multicolor panels
- Fixable and non-fixable options
The Attune Flow Cytometers are high-performance instruments with features, capabilities and specifications that support the demands of researchers at the frontier of immuno-oncology research in the fight against cancer.
Highlights include:
- Faster acquisition of blood and tumor samples
- Less clogging with difficult and precious samples
- Intuitive software for easier compensation
Learn more at thermofisher.com/attune
Simultaneously detect multiple immune-oncology checkpoint proteins with panels designed to run on the Luminex platform. Quantification of soluble checkpoint molecules is a promising new approach that can also be combined with the assessment of cytokines and chemokines in multiplex immunoassays to support research in the field of immuno-oncology and cancer immunology.
Highlights include:
- Human Immuno-oncology Checkpoint panel (14-plex) for the Luminex platform
- Antibodies for key Immune checkpoint markers
- ELISA assays for Immune checkpoint markers
Efficiently use starting material by measuring up to 80 genes of interest in a single well in a 96- or 384-well plate. Based on the Luminex platform, QuantiGene Plex forgoes RNA purification and follows a streamlined ELISA-like workflow.
Highlights include:
- Direct-from-lysate measurement; no RNA purification required
- Works well with FFPE samples containing crosslinked, degraded RNA
- Multiplex up to 80 genes per well
Cancer research webinar
Presented by: Charles Prussak, Pharm D, PhD, UCSD Moores Cancer Center
Title: The Complex Pharmacology of T-Cell CARs
References:
- Batlevi CL, Matsuki E, Brentjens RJ et al. (2016) Novel immunotherapies in lymphoid malignancies. Nat Rev Clin Oncol 13:25-40.
- Junttila MR, de Sauvage FJ (2013). Influence of tumour micro-environment heterogeneity on therapeutic response. Nature 501:46-354.
- Topalian SL, Drake CG, Pardoll DM (2015) Immune checkpoint blockade: a common denominator approach to cancer therapy. Cancer cell 27(4):450-461.
- Grupp SA, Kalos M, Barrett D et al. (2013) Chimeric antigen receptor–modified T cells for acute lymphoid leukemia. N Eng J Med 368:1506-1518.
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Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.