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NanoCellect Biomedical Aims to Bring Cytometry to Benchtop With Rapid, Microfluidic Cell Sorter


NEW YORK (GenomeWeb) – Fresh off of a $10 million Series B funding round, NanoCellect Biomedical has developed a new cell sorting platform using lasers and piezoelectricity to expedite cell differentiation.  

Current cell sorting platforms require experienced users, and are expensive and large. As such, cell sorting equipment is normally limited to a single lab within a massive research institution, and is used to sort a variety of cells for a multitude of researchers. This leads to instrument inavailability, scheduling conflicts, and sterility issues.

In order to address these cost and logistical issues, NanoCellect plans to simplify sample prep using its Wolf cell sorter and N1 single cell dispenser, a compact, robust, portable platform for benchtop or hood use.

As the cells move through the single-cell microfluidic system, it shines a 488-nm laser at the solution, providing information on both scatter and cell fluorescence. Silicon detectors and photomultiplier tubes (PMTs) pick up forward- and backscatter signals respectively. The blue laser excite fluorophores, which then emit signals that are spectrally filtered to resolve up to three colors simultaneously using high-performance PMTs.

Downstream of the sensors, the signals are amplified and converted into the digital domain using analog-to-digital converters. Digital signal processors then allow these signals to become interpretable data for cell sorting.

Within the device's closed disposable microfluidic chip, a low-voltage piezoelectric actuator pushes or pulls the labeled cells into collection channels using water pulses, depending on their level of fluorescence. The actuator reduces shear stress on the cells and eliminates aerosols, which can be a biohazard.  An embedded downstream sorting verification gives instant feedback of cell sorting accuracy.

NanoCellect CEO Jose Morachis explained that researchers can use "up to three colors, and five total optical parameters," including the aforementioned forward and backscatter, and the device can "support two different populations [of cell lines] at the same time."

The overall process on the Wolf cell sorter requires less than 45 minutes from solution to cartridge: 10 minutes to set up the machine, 30 minutes to run and collect the cells needed, and only an additional 20 seconds for clean up. The device can run anywhere from 100 microliters to a milliliter worth of solution, quickly processing up to 24 microliters of sample per minute.

Researchers can run two tubes off the end of the Wolf's cartridge into a the N1 single cell dispenser, which allows research to sort and dispense into 96-well or 384-well plates. Because the microfluidic cartridge has two output ports, it is able to sort cell lines twice as fast. In addition, the entire fluid path is disposable, allowing researchers to rapidly alternate between two sample types.

The sterile cartridge and fluidic tubing are designed for single-day use, preventing any introduction of microorganisms into the cell samples. According to Morachis, the Wolf instrument can detect unlabeled cells and particles with an intensity of lower than 1000 molecules of equivalent soluble fluorochorme, which identify how intense a sample's signal is relative to a solution of the same fluorochrome.

NanoCellect CEO William Alaynick explained that a typical research team using the Wolf cell sorter will have a pool of cells they would like to examine, such as embryonic stem cells. Researchers might manipulate the cells to perform CRISPR on them, for instance, but they would not run the the whole mixture uniformly.

"[Researchers] therefore can run cells through our system, and select cells based on fluorescence to indicate that they have undergone the CRISPR process, and then you can put one cell per well, grow those cells up, and test them later to see if they have the edit that you're looking for, " he said.

Using the firm's technology, researchers can enrich an assay overnight and increase the yield of cells that  they're interested in examining further, reducing downstream costs by enabling more data to be produced, Morachis added.

In the space of personal cell sorting technology, companies such as Becton Dickinson and BioRad have also launched platforms in the past couple of years. BD's FACSMelody is designed to support cell analysis for research applications using multiple lasers and light-based parameters. BioRad's S3e Cell Sorter is equipped with either one or two lasers and up to four fluorescence detectors, in addition to forward and side-scatter detectors. Milliporesigma has also developed its Guava easyCyte flow cytometer that provides up to 14 detection chambers.

"[Some of] these systems are much larger and complex, and come with the maintenance and biohazard issues of high pressure droplet sorters," Alaynick argued. "These systems [also] cannot be used in a standard tissue culture hood."

Founded by a multidisciplinary team of biologist and engineers, NanoCellect spun out of the University of California, San Diego in 2009. Since then, the firm has received $10 million in National Institutes of Health grants for its cell-sorting technology and an additional undisclosed amount of funding through Series A/B financing rounds.  

NanoCellect launched the Wolf cell sorter last winter and collaborated with early-access partners on the product, leading up to the system's official release for research use only in June. In addition, the firm began selling the N1 single cell dispenser in October. Initial prices for the Wolf start at $85,000. While NanoCellect has not published a separate price for the N1 accessory module, Alaynick estimates that the total price, when ordered as a bundle with Wolf, will run the end user about $100,000.

While NanoCellect did not disclose commercial partnerships with downstream companies, the company's website uses 10x Genomics and Illumina platforms as examples for potential downstream sequencing.

"We don't have official partnerships, but we definitely help each other out by working with Illumina's customers, in the sense of complementary technology for their customers," said Morachis.

Emphasizing the product's flexibility and ease of use, Alaynick explained that early adopters have run a variety of sample cell lines on the Wolf system, including stem cells, cancer cells, yeast, bacteria, and cell nuclei.

Ferenc Boldog, director of cell line development at Atum (formerly DNA2.0), mainly uses Nanocellect's Wolf platform to sort strains of chinese hamster ovary cells as well as Horizon Discovery' knockout cell lines. Commenting on the product's limitations that some researchers may face, Boldog believes that the Wolf platform's speed and small scale may inhibit end users from rapidly producing extremely high volumes of cells in a short period of time.

"Frankly, the machine is very user friendly and simple to operate with a minimal footprint, since you don't need an expert to run it," he said in an interview. "The price is also an important advantage, as what you can produce for the price is very attractive."

In an email, Alaynick admitted that developing a sterile, disposable microfluidics path with aerosol-free gentle sorting does have some drawbacks. Using a cartridge requires researchers to "perform backscatter [detection] as opposed to conventional side-scattering". In addition, he noted, microfluidic sorters do not have the same throughput as droplet-based sorters.

NanoCellect envisions its technology used as part of flow cytometry's transition into the "modern age" of microfluidics, improving on the Wolf cell sorter with follow-up products that will target genomic sample prep, single-cell genomics, cell line development, and CRISPR cloning markets.  

"We want to see the company have a major role not only in the flow cytometry space, but also in new and growing applications and tools, like in the CRISPR workflows and immunotherapies," Alaynick added.