The market offers a wide range of excitation wavelengths, equally wide-ranging accurate and sensitive detectors, plus easy-to-use features ranging from validated assays and reagents to simple operation and analysis.
Flow cytometry’s informative potential has been underestimated for many years because of a lack of adequate instruments, automation, reagents, and know-how to approach, integrate, and also substitute other techniques giving single information per assay. In the last decade, flow cytometers have become capable of performing high-throughput screening and high content analysis, evaluating tens of different samples’ features in a single run up to 1536 formats on multiple cell populations. The introduction of imaging flow cytometry has filled the gap between flow cytometry and conventional high content imaging screening, putting flow cytometry at the center of many laboratories, which can now cover with a single instrument the vast majority of needs. The flow cytometry community is a multidisciplinary and diversified group with many different interests and fields of action. These characteristics have prompted the evolution of the techniques, applications, and instruments that allow the use of complex, sophisticated, and standardized and reliable flow cytometric assays.
The world of flow cytometry is now rapidly evolving. During the past few years, there have been significant changes regarding the assays with revived clinical applications, more elegant fluorochrome development, and the size and sensitivity of instruments. There have also been many developments in reagents, instruments, and applications. Polymer chemistry has developed several fluorochromes. For instance, a new fluorochrome strategy constructs large organic molecules into a cage bound by platinum molecules. Specific molecular assembly strategies tune the resonance frequency and fluorescence emission of the cage. Unlike most fluorochromes used today, these structures do not photobleach. The development of more sensitive fluorescence detectors has made smaller instrument footprints possible. This change has been occurring through the developmental history of flow cytometry instrumentation. There are clinical flow cytometry models that have the footprint of a standard laser printer. In the clinical laboratory, these smaller instruments usually have fewer lasers and fluorescence detectors.
The global flow cytometry market will likely rise at a healthy CAGR of 11 percent from 2018–2025, predicts Transparency Market Research. Rising at this pace, the market’s estimated valuation would reach USD 8100 million by 2025. The segment of flow cytometry instruments accounts for a leading market share and in the near future too is expected to hold on to it owing the usage of advanced models for target-based drug discovery. Advancements in the technology such as, the use of more robust photodetectors and new laser emitters, the use of LED lamps as emitter, and changes in analytical capacity in flow cytometry are expected to favor growth of the flow cytometry market. One difficulty staring at the market, on the other hand, is the emergence and soaring popularity of substitutes such as image and scanning cytometers. Limited awareness about the product is also having a negative impact.
Despite half a century of development, there is no sign of flow-cytometry platforms slowing down in innovations. In September 2017, Beckman Coulter revealed some of the advances underway when it released the CytoFLEX LX, which covers a wide range of excitation wavelengths, from the near ultraviolet (375 nm) to infrared (808 nm). For collecting information, it uses avalanche photodiode detectors that provide high and stable quantum efficiency from 400 to 1100 nm. Despite providing so much selection, this platform just sits on a benchtop—not taking up a lot of room in a lab. And scientists get a range of options in selecting the right platform, and then quickly using it. Plus, putting flow cytometry on a benchtop can be done with various platforms. For example, the NovoCyte Quanteon (launched in May 2018 by ACEA Biosciences) sits on a bench, but small size does not mean low power. The NovoCyte Quanteon employs a series of silicon photomultipliers (SiPM), which have superb detection sensitivity. This high-performance instrument is capable of detecting up to 27 parameters with enhanced resolution and walk-away automation capabilities.
Other manufacturers are also providing flow-cytometry platforms with broad capabilities. As an example, the 2100 Bioanalyzer from Agilent can be used with DNA, RNA, or proteins to measure size and quantify and determine purity of a sample. In most flow-cytometry experiments, scientists use one or more fluorophores to label cells or targets. That is easy enough with one fluorophore, but not so easy with a few. So, Thermo Fisher Scientific created its fluorescence SpectraViewer. With this online tool, scientists can test the compatibility of combining specific fluorophores. That information can be used in designing an experiment, rather than finding out afterward that the fluorophore signals overlapped too much for accurate measurements or detection.
These examples already give some idea of what the market currently offers, and—more important—what buyers should expect. That can include a wide range of excitation wavelengths, equally wide-ranging detectors that are accurate and sensitive, plus ease-of-use features that range from validated assays and reagents to simple operation and analysis.
One option that some buyers want with flow cytometry is imaging, which is like combining the capabilities of traditional flow cytometry with a microscope. One of the main advantages of adding imaging is the resulting increase in data. Instead of just counting, sorting, or measuring cells, scientists can look at them. That opens the door for many more analytical options. By virtue of high information content in images, imaging flow cytometry enables to perform more multiparametric analysis of heterogeneous cell populations than conventional non-imaging flow cytometry.
That is not the only technology that manufacturers are combining with flow cytometry. Where automation really matters, manufacturers have put together a collection of platforms to create an advanced workflow. The most important option in flow cytometers is the expanding selection that scientists can consider. In April 2018, for instance, MilliporeSigma unveiled its CellStream flow cytometry system, which uses a camera for detection. It can use as many as seven lasers to analyze cells and submicron particles.
So, even at 50 years old, the flow-cytometry market seems poised to speed up, not slow down. It is not just the improvements in excitation and detection options that matter, but also the improvements in add-on features, such as imaging and automation. As the ability to collect larger volumes and different types of data increases, flow cytometers will give more options for exploring big data and the correlations between it. As a result, the bioinformatics side must keep pace with the hardware advances, because there is no slowing in today’s flowing—not when it comes to analyzing cells and particles sweeping by advanced detectors.
Flow Cytometry As An Emerging Tool For Personalized Medicine
Among the different diagnostic tools available till date, flow cytometry (FCM) offers more precise information at the cellular and molecular level thus emerging as the most prominent technology in diagnosing hematological and oncological diseases. Apart from nucleic acid and cell cycle analyses in cancer cells, rare circulating tumor cells can be easily identified by FCM thus facilitating diagnosis and treatment of hematologic malignancy, including minimal residual disease. FCM offers new avenues in assessing cellular functionality through examination of intracellular compartments with the help of monoclonal antibodies and fluorescence and laser technology. High-throughput quantitative analysis, advancements of in vivo FCM, and assessment of minimal residual diseases, facilitate patient stratification and prediction of leukemia therapeutic response, thus making FCM indispensable in medicine.
FCM is emerging as an important tool in personalized medicine. Inter-patient variability in immune and chemotherapeutic cytotoxic responses is likely due to complex genetic differences and is difficult to ascertain in humans. Cellular phenotyping using FCM identifies cell lines of variable sensitivity to chemotherapeutic agents and aims to identify robust, replicable endpoints of cellular response to drugs that provide the starting point for identifying candidate genes and cellular toxicity pathways for future validation in human studies.
Recent advances in FCM and especially in multi-parameter flow cytometry (MP-FCM) provide the opportunity to obtain high-speed information at real time on damage at the single-cell level. The MP-FCM methodology is based on individual and simultaneous staining of microbial cells employed to investigate their physiological state following different physical and chemical antimicrobial treatments. This facilitates elucidation of the antimicrobial mechanism of action of a given antimicrobial treatment or compound. The combination of MP-FCM methodologies with other conventional methods is namely a promising and increasingly used approach to give further insight in differences in microbial sub-population evolutions in response to antimicrobial treatments. This approach can address the issues related to multidrug resistance of different infectious agents.
Dr Shaik Mohammad Naushad
Head, Biochemical Genetics & Pharmacogenomics,
Flow Cytometry In Cancer Diagnostics
Flow cytometer was first invented by Wallace Coulter in 1953 and the first fluorescence-based flow cytometry device was developed and marketed by Wolfgang Göhde in 1968. After that several laboratories have started using this instrument for clinical diagnosis of cancers and other diseases. Due to advancement of several technological components of flow cytometer such as quality of fluorochromes, monoclonal antibodies, and improved gating strategies, it has become a powerful tool for immunophenotyping of hematopoietic and lymphoid malignancies and therefore it is considered as a future of cancer pathology. Recent studies have also demonstrated that flow cytometry can be used for molecular profiling of breast, colon, and ovarian cancer by evaluating the presence of ER, PR, HER-2/neu, epidermal growth factor receptor (EGFR), e-cadherin, RAS, ERK proteins which are involved in cancer development.
The major factors responsible for the growth of the flow cytometry market are mainly due to the successive technological advancements in the field of flow cytometer as well as increasing incidence of various diseases such as cancer, infectious diseases. Academic and research institutes as well as diagnostic centers are the major end users of flow cytometry and its services are all over the world. The worldwide forecast for the flow cytometry market is expected to increase from USD 3.29 billion in 2017 to USD 4.79 billion by 2022. A Beckman Coulter Life science has recently introduced a highly sensitive and specific CytoFLEX flow cytometer. Besides this Bio-Rad and some of the Indian companies (System India Pvt Ltd.) have also introduced flow cytometers that are highly sensitive and specific for clinical diagnostic purposes. Overall, it appears that flow cytometry is an integral part of diagnostic pathology of cancer and other diseases.
Dr Pravin D Potdar
Indian Dental Association Research Fellowship Program for Year 2017-18