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Flow Cytometry Basics: Benefits, Applications, and Advances

April 26, 2022
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Flow cytometry integrates principles from electronics, fluidics, and optics to measure a variety of physical characteristics of a particle, typically a cell or cellular component. Since its initial design in 1968, flow cytometry has been an integral analytical tool in the life science industry.

Advances in design and performance have led to a rapidly expanding global flow cytometry market, which consists of sales of flow cytometry instruments, chemicals, software, and related services. This market is expected to reach $6.07 billion by 2025 at a compound annual growth rate of 7.6%.1 Next generation flow cytometers break through research barriers, enabling investigation in a variety of new and exciting applications. Let’s take a closer look at the basics of flow cytometry and discover some of its benefits, applications, and trends.

What is flow cytometry? 

Flow cytometry is one of the key detection technologies for measuring physical and chemical characteristics of biological cells. In flow cytometry, sample suspension containing cells or particles is injected precisely into the flow cytometer, and the fluorescent or scattering signals of cells or particles are recorded. The applications of flow cytometry exist in a lot of stages or phases of biology discovery, drug screening, diagnosis, and therapeutic methods, etc. Thus, it has been widely utilized in pharmaceutical and biotechnology companies as well as in research institutes and hospitals.

How Flow Cytometry Works?

Suspended particles in a flowing fluid are analyzed using an optical to electronic coupling system. This system typically relies on hydrodynamic focusing to produce a single stream of particles, which are then subjected to a beam of light to elicit a particles’ unique light scattering signature and emitting fluorescence. Signals are read by detectors, such as photodiodes and photomultiplier tubes, and then converted into electronic signals to be analyzed by a computer. Electrostatic cell sorting can then be utilized to separate the cells, or particles, identified by flow cytometry.

Fluorescent reagents that are utilized in flow cytometry include DNA binding dyes, ion indicator dyes, conjugated antibodies, and fluorescent expression proteins. The properties typically analyzed are a particle’s size, count, complexity, and fluorescence intensity.2

Benefits of Flow Cytometry

Flow cytometry provides analysis capabilities of single cells to large numbers of cells, enabling a robust range of scientific investigations. Fluorescence-activated cell-sorting (FACS) is perhaps one of the best flow cytometry methods to sort heterogeneous mixtures of cells, offering numerous benefits in the fields of immunology, cancer and cell biology. In addition, simultaneous analysis of multiple parameters, quantification of fluorescence intensities, high-speed analyses, and portability further enhance its utility.

Applications

 Immuno-oncology and Infectious Diseases

 Flow cytometry has been utilized in applications such as immunology and cancer biology, as well as infectious disease monitoring. Recently, flow cytometry has played an integral role in COVID-19 pandemic. Enabling researchers to perform rapid cellular and particle analyses for diagnostic, vaccine, and therapeutic development.

There have also been significant advances in applications such as the immune system and its response to cancer. Flow cytometry stands out as a key tool to enable simultaneous characterization of mixed populations of cells from blood and bone marrow. Additionally, it optimizes solid tissue analysis by dissociating samples into single cells such as lymph nodes, mucosal tissues, and tumors. 4

Environmental and Wastewater

Flow cytometry is an excellent resource for bacterial health assessments in the wastewater treatment industry. Biological wastewater treatment relies on healthy bacteria populations to breakdown waste. Flow Cytometry provides key analyses on the bacterial health of a given system, giving operators and environmental staff fast and reliable data to manage their facilities.5

Microbiology and Metabolic Diseases

Apoptosis, or programmed cell death, is a significant mechanism of interest in a variety of fields including metabolic diseases, cell biology and cancer. Flow cytometry provides researchers with a great tool for real-time apoptosis detection.

Advances and Trends

High-Throughput Flow Cytometry

High-Throughput Flow Cytometry is an emerging technique that utilizes high-speed cell parameter processing to analyze several heterogenous cell groups simultaneously.

Imaging Enhanced Flow Cytometry

High speed camera enhanced imaging enables researchers to investigate new applications by merging cell morphology imaging with size, granularity, and multiplexed protein expression data.

Acoustic Focusing

Acoustic focusing utilizes radiation pressure forces in leu of, and in addition to, hydrodynamic focusing to position particles or cells for analysis in the flowing stream. When hydrodynamic and acoustic focusing are utilized together, high precision analyses with broader dynamic ranges of volumetric sample input rates are possible.

Outsourcing, and Integrated, Embedded Teammates

Flow cytometry needs are typically handled on-premise in large pharmaceutical labs, but there has been a shift over the past decade where labs are outsourcing some of these requirements. Outsourcing has proven valuable for certain studies, but ultimately, having expertise on-site, close to the research teams, is the optimal environment for critical milestones to be hit.

Not only does this reduce logistics costs for sending samples out, but it drastically reduces turnaround time to get results. An integrated service team managing flow cytometry needs, brings expert services inside the customer labs.

Conclusion

The high-speed, multi-parameter capabilities of next generation flow cytometers have positioned this technology as a leading biomedical instrument for applications related to immunology, cell biology and environmental science. Advances in imaging, acoustic focusing, and high-throughput capabilities add even greater value to this line of instrumentation, driving its projected market expansion in the next few years. Additionally, flow cytometry has rapidly become a cornerstone technology in the development of rapid vaccines, diagnostics, and therapeutics in response to the Covid-19 pandemic. The growing global threat of future pandemics will no doubt rely on flow cytometry and its continued development.

With the advancement of applications and more complex technology, flow cytometers are becoming increasingly more difficult for the average user to unlock their full potential. Only dedicated specialists, with years of experience, can ensure proper instrument support and data integrity. Service providers support these specialists with standardized procedures and access to a greater network of flow subject matter experts.

Click here to discover how PerkinElmer Flow Cytometry Instrument Concierge Services can help you and your research team maximize the usage of your flow cytometry instrument fleet.

References:

  1. “Flow Cytometry Global Market Report 2022.” Benzinga, www.benzinga.com/pressreleases/22/03/n25995316/flow-cytometry-global-market-report-2022. Accessed 28 Mar. 2022.
  2. Introduction to Flow Cytometry: A Learning Guide Introduction to Flow Cytometry: A Learning Guide. 2002. BD-Flow-Cytom-Learning-Guide.pdf (bu.edu)
  3. Gil-Manso, Sergio, et al. “Comprehensive Flow Cytometry Profiling of the Immune System in COVID-19 Convalescent Individuals.” Frontiers in Immunology, vol. 12, 6 Jan. 2022, p. 793142, www.ncbi.nlm.nih.gov/pmc/articles/PMC8771913/, 10.3389/fimmu.2021.793142. Accessed 28 Mar. 2022.
  4. McKinnon, Katherine M. “Flow Cytometry: An Overview.” Current Protocols in Immunology, vol. 120, no. 1, 2018, pp. 5.1.1–5.1.11, 10.1002/cpim.40.
  5. “Benefits of Flow Cytometry in Wastewater.” Environmental Business Specialists, 10 Dec. 2021, www.ebsbiowizard.com/articles/benefits-of-flow-cytometry-in-wastewater/. Accessed 28 Mar. 2022.

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