Cell sorter, dispenser for gene editing

Gene editing technology requires modification, identification, and clonal expansion of a single cell. However, many labs can’t afford the automated flow cytometry and sorting operations necessary for gene editing due to the cost of owning and operating an in-house sorter.

A new technology allows specialists to conveniently sort cells in their own labs, on their own equipment, and on their own time. NanoCellect Biomedical’s WOLF Cell Sorter enables users to sort cells with greater confidence that the cells will survive. The Cell Sorter does not use conventional jet-in-air cell sorter technology that can damage sensitive cells such as stem cells by using high pressures that can negatively alter their biological behavior. With an on-chip fast valve, the sorting mechanism gently guides cells into collection channels. Sorting takes place inside a microfluidic chip and avoids creating dangerous aerosols emitted by traditional cell sorters. Smarter cell sorting allows greater flexibility or samples that normally require work in a hood.

The Cell Sorter uses five optical parameters for sterile single-cell sorting into 96- and 384-well plates with well-by-well data indexing for clonal expansion. With a footprint less than 2ft³, it can be placed in any hood for sterility. Its rapidly exchangeable fluidics and cartridge eliminate carry-over between samples and provide one-minute clean-up.

Depending on the experimental goals of the lab, characterization of clones can also be performed via flow cytometry (see Figure 1).

After designing, developing, and manufacturing the WOLF Cell Sorter, the company realized users needed a single cell dispensing unit to go with the sorters. Since their equipment is already being used in a sterile environment, all that was required was to extend tubes from the end of the cartridge and into the dispenser. This allowed the dispenser to maintain a gentle, sterile, and microfluidic domain. The entire fluid path, the cartridge, and the dispenser tubing is disposable so the machine can facilitate rapid changeover between sample types.

Motion control design features

To ensure the single-cell dispenser would operate accurately and repeatedly for the WOLF Cell Sorter’s entire life cycle, it needed a robust motion control unit. NanoCellect engineers also needed a motion controller that was easy to install and would integrate quickly with simple software for ease of use.

The N1 Single Cell Dispenser is helpful for people who are doing antibody discovery, cell line development, CRISPR cloning (a technology for gene editing), or working with single cells upstream of demanding applications. Because the microfluidic cartridge has two output ports, users can sort twice as fast as more expensive models on the market.

The engineering team at NanoCellect used AllMotion’s EZ4AXIS Controller to control the positioning system (see Figure. 2, Page 16). The EZ4AXIS, a four-axis position controller with built-in power drivers, only uses three of the controller’s four axes for X, Y, and Z operations. Stepper motors in the device offer extreme accuracy. The dispenser requires very low torque because the load includes only small plastic dispensers. Many applications require the sorting and dispensing of single cells, such as B-cell identification in antibody discovery, and monoclonal Chinese hamster ovary (CHO) cell line development. Furthermore, single-cell omics assays, where sample preparation is a key step for quality data generation, are no longer heroic academic exercises.

The N1 Single Cell Dispenser remains aerosol-free, sterile, and disposable.

Two outlets from the sorting cartridge are attached to tubing and two dispensing heads. Users can select the same cell population for every well or assign two different cell populations as they are sorted from the sample (see Figure. 3). Users can also simply dispense from a pure population.

Positioning control of the dispenser is initialized from the company’s controller software through a serial port string into the AllMotion EZ4AXIS controller board. The controller board provides NanoCellect with ease of interfacing, programming, and quality of operation. Once the controller receives the command string, it sends the precise movement data instructions directly to the motors. Operation is smooth, accurate, and repeatable. The N1 Single Cell Dispenser is integrated into the intuitive software, allowing the user to simply select Single-Cell from the dropdown menu, map out the plate, and hit go.

The EZ4AXIS can simultaneously control multiple movements independently. It is one of the smallest controller/drivers on the market – and one of the most powerful for its size.

AllMotion
https://www.allmotion.com

NanoCellect Biomedical Inc.
https://nanocellect.com