Trending Toward Miniaturized Flow Cytometry

Flow cytometry is currently the gold standard for analysis of cells in the medical laboratory and biomedical research. The traditional techniques of flow cytometry have been the stalwart of cell analysis for many years. But times have changed – scientific breakthroughs, grant applications, and publications wait for no one – and there are now a host of new technologies that complement this traditional modality. With the advent of automation and high-throughput technologies, researchers no longer need to spend hours in a darkroom to capture a simple fluorescent image, painstakingly measuring cell parameters one cell at a time, or risk losing precious samples due to mechanical issues.

Even though flow cytometers provide good sensitivity and impressive throughput of thousands of cells per second, commercial systems are bulky, costly, and require trained personnel for operation and maintenance. This has limited their use to the central laboratory and core facilities. Fueled by the need of point-of-care diagnosis, a significant effort has been made to miniaturize and reduce cost of flow cytometers. Microflow cytometers that combine microfluidics and miniaturized detection systems are a promising solution for future diagnosis. The lab-on-a-chip platform has been used to develop such systems during the last years.

Indian Market Dynamics
The Indian flow cytometers market in 2016 is estimated at Rs. 205 crore, with reagents constituting 60 percent market share. The analyzers and cell sorters market continues to be dominated by BD India and Beckman Coulter.

The government is moving toward centralized facilities, and for any instruments having higher than average unit price of Rs. 25 lakh, tenders are not being finalized at the regional level. Also, with the buying life cycle, especially for mid-end and high-end equipment normally being up to 12 months, some vendors abstain from participating.

Over the last couple of years, HIV spends gave way to cancer immunology where the private sector is more active. The government spent more funds for stem cell research and infectious diseases. Some buying was done by the private sector, for instance, Amrita Medical College, with funding by the government.

Technology Trends
The past decades have marked a number of key technological advances in flow cytometry, all of which increase the number of parameters that can be measured simultaneously from single particles. For conventional flow cytometry, the most notable advancement has been the extension of the alignical design with additional and more powerful lasers. Together with advances in fluorophore design, 18-parameter flow cytometry is now routinely used; 30-parameter flow cytometers have recently become commercially available; and 50-parameter flow cytometry is projected to be available soon.

Spectral flow cytometer. The arrival of the spectral cytometer opens a new era in flow cytometry by allowing detection of the whole spectrum of each particle, from ultraviolet to infrared. It gives a great deal of information and details on the fluorescence emission. The spectral cytometer is able to simultaneously discriminate between the spectrums without them overlapping, capable of using up to 32 different parameters, and distinguishes the shapes of emission spectra along a large range of continuous wave lengths.

CyTOF. Cytometry by time-of-flight (CyTOF) is a new method for detecting antibodies bound to cells. CyTOF overcomes the loss of resolution occurred from auto fluorescence and spectral spillover of conventional flow cytometry. Use of traditional labeling techniques with minimal change to current protocols, easy panel design, minimal to no compensation need, and no auto-fluorescence are some of the advantages offered by this new technology.

IFC. Imaging flow cytometry (IFC) combines the single-cell imaging capabilities of microscopy with the high-throughput capabilities for conventional flow cytometry. Recent advances in IFC are remarkably revolutionizing single-cell analysis and have fil ed in the gaps to provide both quantitative power of flow cytometry and fluorescence localization power of imaging in a single platform. It combines flow cytometry with a high-resolution multispectral imaging system, acquiring several images per cell, at current rates of 5000 cells per second.

Optofluidic microflow cytometers. They are integrated with optics to provide significant enhancements for flow cytometry. Low costs, higher sensitivity, free optical alignment, and smooth interaction interfaces are the obvious advantages. Owing to the recent development of lab-on-chip (LOC) technology, microfabrication, and micromachining techniques, the miniaturization of a flow cytometer can be achieved. These cytometers offer significantly lower costs and size reductions, as well as low reagent requirements and portability advantages over a bench-top flow cytometer.

Acoustic focusing. Modern flow cytometers handle thousands of events per second. However, when increasing the flow rate, one has to lower the overall resolution. Scientists have solved this issue with acoustic-assist focusing. They use sound energy to usher the particles into the center of the sheath fluid, where hydrodynamic focusing takes place. This allows high resolution at a high flow rate. In addition, this technology also enables the possibility of running multiple streams at the same time in one flow cytometer.

Way Forward
Flow cytometry will grow in importance to become a USD 6.3 billion industry by the end of the decade. The market is set to grow at a compound annual rate of more than 9 percent through 2020, although that expansion will be muted somewhat by the generally high cost of equipment. Small-size high-throughput cytometers are expected to gain popularity over the coming years due to associated benefits such as ease in use and cost-effectiveness. Furthermore, improvement in fluorescent dyes and introduction of bench top cytometers are the other growth propellers. For instance, multicolor flow cytometry coupled with multiple lasers is the fastest growing

application segment, which finds extensive applications in the field of R&D innovations in new drug development and is adopted by many contract research organizations.

The industry is expected to witness significant changes over the next five years as the technology establishes a growing base in biotechnology labs for applications like counting, sorting, biomarker detection, and protein engineering.

The expansion of flow cytometry applications from cellular analysis to molecular and genomic analysis, increasing research and development spending, and a shift in small-molecule drug discovery will grow the market. Recent market consolidation, a trend that is expected to continue over the next 5 years, has already produced strong, vertically integrated competitors that are better able to compete as sole-source vendors to end users.