Flow Cytometry Applications: Advanced Uses, Benefits, and Limitations

October 17, 2024

Damien Montamat-Sicotte, Scientific Business Director

Flow cytometry is a technique used to measure the physical and chemical characteristics of cells or particles as they flow in a fluid stream through a beam of light. By labeling cells with fluorescent markers, flow cytometry can analyze multiple parameters simultaneously, providing a comprehensive profile of each cell.

This guide explores flow cytometry applications and various uses, providing insights into its significance in clinical research.

About Flow Cytometry and Its Impact

Flow cytometry integrates principles and techniques from electronics, fluidics, and optics. Samples stained with fluorescent labelled antibodies are suspended in fluid and injected into the flow cytometer.

As cells pass through a laser beam, they scatter light and emit fluorescence that is detected and measured. This data provides detailed information about the cell’s properties, including size, complexity, and the presence of specific markers, allowing to determine a cell’s phenotype and confirm protein expression either extra- or intra-cellular.

Flow Cytometry in Research and Clinical Settings

Flow cytometry offers several advantages, including:

• High-throughput single-cell analysis: Ability to analyze thousands of cells per second.
• Multiparametric data: Simultaneous measurement of multiple parameters and confirm protein expression.
• Precision and sensitivity: Detection of rare cell populations.

These features make flow cytometry an essential tool in various fields, from basic research to therapeutic monitoring.

For example, in immunology, it can be used to analyze the distribution and characteristics of different immune cell types, aiding in the understanding of immune responses and potential therapeutic targets.

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Flow Cytometry platforms and services by CellCarta

10 Key Applications and Uses of Flow Cytometry

1- Clinical Monitoring

Hematological Malignancies:

Flow cytometry is instrumental in monitoring blood cancers such as leukemia and lymphoma. It identifies abnormal cells based on their surface markers, enabling precise disease classification and monitoring.

For example, the flow cytometry platform allows for the measurement of minimal residual disease (MRD) in leukemia patients, helping understand treatment responses.

Immunophenotyping:

This application identifies and quantifies different immune cell populations, aiding in the phenotyping of immune disorders. Flow cytometry can detect specific markers on the surface of immune cells, providing detailed information on immune status.

CellCarta’s platform agnostic expertise in flow cytometry, whether conventional, mass or spectral, allows for immunophenotyping tailored to the project’s need.

Basophil Activation Test (BAT):

The Basophil Activation Test (BAT) is frequently employed to measure allergy severity and assess clinical responses to immunomodulatory treatments. Flow cytometry enable the precise monitoring of the basophil population by analysing degranulation markers such as CD63.

2- Cell Sorting and Cell Isolation

Flow cytometers equipped with sorting capabilities can physically separate specific cell populations from a mixed sample. This is crucial for research applications requiring pure cell populations, such as stem cell research or cancer studies.

For example, CellCarta‘s advanced flow cytometry techniques facilitate the isolation of rare cell populations for downstream, phenotypic, functional, genomic and proteomic analyses.

3- Cell Cycle Analysis- Assessing Cell Replication States

Flow cytometry analyzes cell cycle phases, helping researchers understand cell proliferation and apoptosis.

This is particularly important in cancer research, where cell cycle dysregulation is a hallmark of disease. By measuring DNA content and identifying different phases of the cell cycle, researchers can evaluate the effects of anticancer drugs and understand tumor growth dynamics.

Cell proliferation assays to measuring Cellular Metabolic Activity: These assays evaluate cell division and growth in response to various stimuli. Flow cytometry tracks cell proliferation by labelling cells with a fluorescent dye, providing insights into cellular responses and drug efficacy.

4- Apoptosis and Necrosis Detection

Flow cytometry distinguishes between apoptosis (programmed cell death) and necrosis (cell death due to injury).

This is vital for studying cellular responses to treatments and understanding disease mechanisms.

For example, in drug development, determining whether a compound induces apoptosis or necrosis can inform its potential therapeutic value.

5- Immune System Monitoring

Tracking Immune Cell Populations:

Flow cytometry monitors changes in immune cell populations, such as:

• CD4 and CD8 T cells, in patients with HIV.
• Naïve and memory B cells in auto-immune patients following treatment and reset of their B cell populations (immune reset hypothesis).

This information is crucial for assessing disease progression and treatment efficacy. Regular monitoring helps clinicians adjust therapies to maintain immune function and manage the disease effectively.

6- Cancer Research- Analyzing Tumor-Infiltrating Lymphocytes (TILs)

Flow cytometry examines immune cells within the tumor microenvironment, aiding in immunotherapy research.

This application helps identify how immune cells interact with cancer cells, providing insights into tumor immunity and helping develop strategies to enhance immune responses against tumors.

7- Blood and Platelet Analysis

Evaluating Platelet Function: Flow cytometry assesses platelet activity and function, important for diagnosing and managing clotting disorders.

This technology provides detailed information on platelet count, activation, and aggregation, aiding in the diagnosis of conditions like thrombocytopenia and platelet function disorders.

8- Stem Cell Research

Identifying and Sorting Stem Cells: Flow cytometry is essential to identify and isolate stem cells based on specific surface markers.

This application is crucial for regenerative medicine and stem cell therapy research, where pure populations of stem cells are needed for developing treatments for various conditions, including neurodegenerative diseases and tissue injuries.

9- Microbiology and Virology

Pathogen Detection: Flow cytometry detects and quantifies bacteria, viruses, and other pathogens in clinical samples. This is vital for infectious disease research and diagnostics, providing rapid and accurate pathogen identification.

For instance, it can be used to monitor bacterial contamination in water supplies or to detect viral infections in patient samples.

10- Drug Development and Toxicology

Flow cytometry assesses how drugs impact cell populations, viability, and function. This application is crucial for drug discovery and safety assessments, helping to screen potential therapeutic agents for efficacy and toxicity.

By measuring parameters such as cell viability, apoptosis, and proliferation, researchers can determine the effects of new drugs on different cell types.

Using specifically antibodies against intra-cellular phosphorylated biomarkers (phosphoflow) allows the monitoring of signaling pathways. With the profiling of key signaling pathways using phosphoflow, drug developer get a better understanding of their compound’s mechanism of action.

Explore all our Flow Cytometry Services and Platform

Considerations when developing Flow Cytometry projects

Flow cytometry’s ability to provide detailed analysis of cell populations makes it indispensable in clinical research. With the power to confirm protein expression in specific cell types at a single-cell level, its applications span various fields.

Flow cytometry has revolutionized many aspects of biomedical research and clinical practice. Its applications in immunology, oncology, microbiology, and infectious diseases have led to significant drug discoveries and innovations. For example, flow cytometry has played a crucial role in the development of immunotherapies for cancer, enabling researchers to understand how immune cells interact with tumors and identify new therapeutic targets.

Flow cytometry is a versatile technology in modern science that offers numerous applications in clinical research and therapeutic monitoring. Here we provided a quick highlight of about 10 key applications and uses with helpful insights and links to our Flow Cytometry services.

Learn about CellCarta and all our solutions.

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