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Lightcast Discovery Developing Droplet-Based Single-Cell Screening Platform

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NEW YORK – Single-cell technology company Lightcast Discovery is developing a droplet-based platform that promises to enable high-throughput single-cell functional assays in a massively parallel fashion.

The Cambridge, UK-based company raised £38 million ($49 million) in Series B funding last year and recently opened an office and laboratory in the US.

Founded in 2019, Lightcast has been developing a droplet-based microfluidic platform that harnesses so-called optical electrowetting-on-dielectric (oEWOD) technology, according to Paul Steinberg, the firm's chief commercial officer.

The company acquired its founding IP for oEWOD from Base4 Innovation, another Cambridge, UK-based company that was developing a microdroplet-based DNA sequencing platform and has since been dissolved, and has been expanding its IP portfolio over the last several years, Steinberg said.

While Cameron Frayling, Base4's founder and CEO, lists himself as the founder of Lightcast on his LinkedIn profile, Steinberg clarified that Frayling has not been directly involved with the company since its inception.

Lightcast is developing the oEWOD technology to perform "highly controlled" sequential and multiplexed single-cell assays to achieve complex cell profiling under the control of light, according to the company.

As part of the workflow, picoliter-scale droplets, which contain reagents or analytes, such as cells or beads, are generated using low-shear-stress droplet generators. Steinberg said the technology can currently accommodate up to eight different soluble reagents or objects in one workflow, which are loaded into droplets sequentially using separate fluidic chips.

Once generated, the droplets are loaded onto the temperature-controlled oEWOD microfluidic chip under pressure, where they are held between two layers of transparent substrates. One substrate contains a photoactive layer, and its local electric field can be changed by the application of light. This changes the local wetting that holds the droplets and allows droplets to be moved by individual points of light, termed sprites.

Under the control of assigned sprites, droplets are first moved across a detection window where they are screened by a camera for their size, occupancy, and content, and any unwanted or unoccupied droplets are removed.

The remaining droplets are put into arrays, where they can be merged to carry out simultaneous single-cell assays. These generate readouts that can be analyzed across the array in parallel. Steinberg said the company's platform currently has two detection modalities: brightfield imaging, which can visually query individual droplets, and fluorescence detection, which enables fluorescent-based cellular assays at the single-cell level.

In the end, single cells can also be dispensed onto well plates for further downstream analysis, such as nucleic acid sequencing.

At launch, the company's platform will be able to manipulate tens of thousands of droplets in parallel, Steinberg said, and the team already has a roadmap to further scale throughput to hundreds of thousands of cells.

Besides the hardware, which will be a benchtop instrument, the company also plans to launch a series of applications with optimized reagent kits and protocols.

"Our aim is developing the platform to be as powerful as possible, which is not just the hardware itself but all of the reagents, the protocols that would be required [for the workflow], to be as push-button friendly as possible for our customers," Steinberg said.

He declined to disclose specific applications Lightcast plans to release first but said that the company is broadly interested in areas including antibody discovery, cell and gene therapy, and drug development. Its target customers will primarily be working in translational research as well as in therapeutics development, he added.

The company kicked off an early-access program last year, he said, to engage with experts in the translational research and drug development fields to help refine and enhance its technology and workflows. He declined to comment on how many collaborators the program has signed on so far but noted it is "still pursuing additional partnerships."

Currently, Lightcast has about 110 employees, working primarily at its Cambridge headquarters. As the company heads toward product launch, it will expand its commercial team, Steinberg said. The firm raised $49 million in Series B funding last year for the development and commercialization of the platform, he added.

Lightcast is not the only company eyeing the massively parallel single-cell functional analysis market. Waltham, Massachusetts-based Flexomics, for instance, is also developing a platform capable of simultaneous functional and genomic analyses of hundreds of thousands of individual cells in parallel. That firm's approach combines live-cell analysis and single-cell genomics, enabling the interrogation of cellular interactions and dynamic responses.

This is also not the first attempt to harness oEWOD as a research tool, as Base4 had previously tried to commercialize the technology for DNA sequencing. While Steinberg said he is confident that Lightcast will bring the technology to the market soon, he declined to disclose a specific timeline. "We are well down the road of our commercialization path, so it is not years away," he said.