Applications of Modern Clinical Flow Cytometry

Current applications of flow cytometry in the clinical laboratory

24 Mar 2016
Sonia Nicholas
Managing Editor and Clinical Lead

Editorial article

Flow cytometry measures optical and fluorescence characteristics of individual cells as they flow, in single file, through a stream of liquid

Flow Cytometry Principle Flow cytometry measures optical and fluorescence characteristics of individual cells as they flow, in single file, through a stream of liquid. As each cell flows through the liquid stream, it passes by a laser. The cell’s size, internal complexity, DNA/RNA and protein content can be determined by measuring the light scatter produced.

Flow cytometry, now well established in many hematology and immunology labs, is a technology that has had a substantial impact on the monitoring and diagnosis of disease over the last decade.

Current applications

In hematology, immunophenotyping is used to study proteins expressed by cells, so that they can be accurately identified. Immunophenotyping by flow cytometry is used for the diagnosis of new leukemias, it allows certain acute myeloid leukemias to be distinguished from one another, and it can also be used to detect minimal residual disease.

Flow cytometry can be used to identify platelet abnormalities such as autoimmune thrombocytopenia purpura and those associated with conditions such as Bernard-Soulier disease. Flow cytometry can also aid diagnosis of red cell disorders such as hereditary spherocytosis and paroxysmal nocturnal hemoglobinuria.

Immunological flow cytometry is used for HIV diagnosis as well as viral load monitoring, it is also used in the diagnosis of other immunodeficiences and allergy testing. Flow is also used by blood banks for the identification and quantitation of fetal blood cells in maternal circulation following a feto-maternal haemorrhage. Flow cytometry can also be used to detect red cell bound immunoglobulins and red cell antigens, which is especially useful in patients who have received multiple transfusions. The technology can also be used to assess leukocyte contamination of leukocyte-reduced blood products.

Technology advances

FC 500 Series Flow Cytometer, Beckman Coulter

Flow technology has advanced considerably over the last decade. Bench-top flow cytometers are now equipped with 2 or 3 lasers and are able to analyze multiple samples in a relatively short space of time. Sophisticated hardware and software enables flow cytometrists to spend less time on sample prep and analysis and more time assessing results and providing valuable interpretive reports. You can find the latest technical information on flow cytometry in our download library and product directory.

Available Flow Cytometry downloads from the SelectScience® library:

Multiparametric Flow Cytometric Evaluation of Cytotoxic T Lymphocyte Populations

No-Lyse, No-Wash Protocol for 6-Color Immunophenotyping Analysis of Stained Human Whole Blood without Compensation Using the Attune NxT Flow Cytometer

Immunophenotypic Analysis of Human Blood Leukocyte Subsets Using a 13-Color Antibody Panel Using the NovoCyte Sytem

6-Color Panel for Staining Human Peripheral Blood Lymphocytes

Identification of B Cells, NK Cells, Multiple T Cell Subsets and Myeloid Cells by 13-Parameter Immunophenotyping of Human Lysed Whole Blood

A Novel Imaging Cytometry Method for Immunophenotyping

Looking to purchase a new analyzer? Download our exclusive Flow Cytometers Buying Guide

View the flow cytometer product directory

NovoCyte Flow Cytometer

Agilent Technologies

NovoCyte is a high-performance benchtop flow cytometer designed for all levels of users and all types of laboratories. The budget-friendly instrument can detect up to 17 parameters with enhanced sensitivity and resolution. The customizable laser and optical configurations of the NovoCyte offer a high degree of flexibility while providing complex cell analysis capabilities. The NovoExpress software facilitates easy and intuitive sample acquisition and analysis. Automation of multiple fluidic functions eliminates cumbersome and time-consuming procedures. Manual sample handling is minimized by the flexible NovoSampler Pro. The system can automatically analyze samples in single tubes, multi-tube racks, or 24-, 48-, or 96-well plates

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Invitrogen™ Attune™ NxT Flow Cytometer

Thermo Fisher Scientific

The Invitrogen™ Attune™ NxT flow cytometer can be designed to accommodate the most common fluorophores and fluorescent proteins used in flow cytometry to match the panels you are currently running. Multiple fluorescent proteins can be detected with an optional 561 nm laser. Whether you configure your system now or upgrade later, the Attune NxT flow cytometer can grow with you and your research needs. The Attune NxT flow cytometer, designed using acoustic-assisted hydrodynamic focusing, achieves sample-throughput rates of of 12.5 µL/min–1 mL/min or 12.5–1,000 µL/min—up to 10 times faster than traditional hydrodynamic focusing systems and data acquisition speeds of 35,000 events/sec. Acoustic focusing allows significantly higher sample rates, which simplifies sample preparation, reduces protocol times, and minimizes cell loss. This feature is useful for no wash, no lyse immunophenotyping protocols using whole blood and for samples that are inherently low in concentration, including cerebrospinal fluid and stem cell preparations. Your next result relies on the ability of your experimental design and your instrument to deliver more—more data, more detail, and more throughput. The Attune NxT flow cytometer was designed to meet the demands of more complex experiments, offering reliable data, day after day, in a grow-with-you system.   Features & Benefits: Modular design—Multiple configurations available—field upgradable. Save time—10X faster speeds with no loss in data quality. Simplified sample prep—No wash, no lyse options, non-clogging fluidics. Enables unique applications—Complex protocols on a broad range of cell types and samples.

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Blood TransfusionBlood Transfusion involves giving donor blood to a recipient patient. Screening is essential to avoid transfusion reactions. Blood banks use immunohematological techniques to determine rh and ABO blood group, and screen for antibodies using specific antisera. Blood Banks use Direct Coombs Tests (DCT) and Indirect Coombs Tests (IAT) to detect hemolysis and Haemolytic Disease of the Newborn (HDN).HematologyIn Haematology / Hematology, complete blood cell counts (or full blood counts) are obtained using automated blood count analyzers to enumerate blood cell types.  Hematology also encompasses haemostasis and coagulation, thrombophilia and hemophilia, plasma viscosity and ESR analysis, hemoglobinopathies, cell morphology and haematinic measurement.ImmunologyImmunological techniques measure and characterize immune responses. Immunology kits and analysis systems often use techniques such as ELISA, radioimmunoassay (RIA) and immunodiffusion assays, Immunohistochemistry, and flow cytometry. Immunologists use equipment such as flow Cytometers, plate readers, plate washers and fluorescent microscopes.Point-of-CarePoint-of-Care Testing (POCT) or Near Patient Testing (NPT) products are available for urine, blood and other clinical chemistry analyses. POCT includes: blood glucose testing, blood gas and electrolytes analysis, rapid coagulation testing (PT / INR), rapid cardiac markers diagnostics, drugs of abuse screening, urine strips testing, pregnancy testing, fecal occult blood analysis, food pathogens screening, hemoglobin diagnostics, infectious disease testing and cholesterol screening.Clinical GeneticsMolecular Genetics covers the analysis of hereditary genetic disease and chromosomal abnormalities. Genetics can be analysed using DNA, RNA, and protein microarrays, PCR, RT PCR and DNA sequencing. Genetic equipment includes genetic workstations, thermal cyclers, cooling blocks and electrophoresis products. Diagnostic kits are used for DNA / RNA extraction and purification.CytometryCytometry is a broad term for the quantitative analysis of cells and cell systems. The most popular methods to study cytometry are flow cytometry and image cytometry.LeukemiaImmunophenotypingHIV and AIDSBlood tests are the most common way to identify and monitor HIV and AIDS. Current methods include ELISA, Western blot, RT-PCR viral load and CD4/8 testing. Areas for development include point-of-care assays for use in developing countries, and self-testing ‘at home’ HIV kits.VirusesViruses are microscopic pathogens that require a host cell to replicate. Understanding their structure, replication cycle, and impact on the immune system is crucial in developing effective treatments, vaccines, and diagnostic methods. Research continues to focus on emerging viral diseases and antiviral drug discovery. Explore solutions for virus research, detection, and treatment in our peer-reviewed product directory; compare products, check customer reviews, and get pricing directly from manufacturers.Blood AnalysisThe analysis of blood is vital for many areas of life sciences and forensic investigations. Blood samples can be tested for a number of different reasons such as diagnosis, glucose levels, cholesterol and drug testing.platelets