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

FDA approvals

Med-El USA’s cochlear implant (CI) system is approved for single-sided deafness (SSD) and asymmetric hearing loss (AHL). CI’s Synchrony and recently FDA-approved Synchrony-2 are now indicated for individuals 5 years and older with SSD who have profound sensorineural hearing loss in one ear and normal hearing or mild sensorineural hearing loss in the other ear, or individuals 5 years and older with AHL who have profound sensorineural hearing loss in one ear and mild to moderately severe sensorineural hearing loss in the other ear, with a difference of at least 15dB in pure tone averages between ears. 

Florida Anodize System & Technologies Inc.’s quality management system for its medical device surface coating services was audited by TUV SUD and found to be in full compliance with ISO 13485:2016.

Nanotech Precision, a manufacturer of micro-sized instruments and electrodes, has received ISO 13485:2016 for medical devices and quality management systems.

The MV1200-S Advance has a manual vertical front door, 2-axis linear shaft motor, U-shaped hardened table, and submerged machining to 8.7" depth. Other features of the MV1200-S Advance enhancing small medical part manufacturing include:

Cylindrical drive technology: Round magnetic shaft of linear motor creates 360° magnetic flux for no-contact, smooth, friction-free motion; larger air gap between shaft and forcer minimizes heat output, effect of metallic shop dust on system

Mitsubishi M800 CNC control: Intuitive operation via 19" touch screen with ergonomic tilt/swivel mounting system

Advance pendant box: Thin, tapered design; 3.5” LCD touch screen, provides operators functionality at machine, minimizing setup

Auto-threading system: For small parts production, 14" annealing length

Integrated passive components (IPCs) are attracting increasing interest due to the miniaturization of wireless devices, as well as the need to increase reliability of signal conditioning in radio frequency (RF) circuits such as filtering, impedance matching, differential to single-ended conversion, and coupling.

IPCs are electronic sub-systems that combine multiple discrete passive components into a single surface-mounted device. Manufactured using low temperature cofired ceramic (LTCC) technology that allows passive components to be layered in 3D, IPCs deliver the same functionality as 10 to 40 individual components, reducing required board space.

With this approach, the entire front-end between RF chipset and antenna can be manufactured in a single, ultra-low profile (0.35mm to 1.0mm total thickness) package that is less than 20% the size of the same circuit comprised of discrete components.

“If you were to look at the circuit from above, you would only see three chips: the RF wireless chip, the IPC, and the antenna,” explains Manuel Carmona, RF business development and product manager at Johanson Technology. “The design is clean and small. You can make the overall printed circuit board (PCB) size much smaller using IPCs while maintaining high wireless performance.”

IPCs also deliver greater reliability. By creating a circuit within a small LTCC package, variability and potential points-of-failure are nearly eliminated. A single integrated package of matching components also assures compliance with FCC or ETSI requirements.

“A lot of the variability of solo surface-mounted components is due to the individual variance of each component. When you multiply that by 10 to 20 components, you just need one weak link in the RF chain for the entire front-end to fail,” Carmona says.

IPCs are available for almost any type of passive circuit, including low- and high-pass filters, diplexers, triplexers, impedance matched baluns, balun-filters, band pass filters, and couplers.

LTCC technology

Manufacturing IPCs is similar to creating multi-layer surface mounted device (SMD) component parts, such as capacitors and inductors. However, LTCC manufacturing allows circuits to be embedded in up to 40 separate layers of a 3D package that is still very low profile.

Using this manufacturing process, Johanson Technology developed a line of small, highly reliable IPCs for RF systems. These components operate across several bands from 300MHz to 10GHz, covering cellular, DECT, WLAN, Bluetooth, 802.11 (a, b, and g), and GPS applications.

Each integrated package is RF tested to ensure all components are working properly and are fully integrated.

“Because the LTCC manufacturing process is extremely precise and repeatable, we can stay within the manufacturing limits of alignment, variations, and so forth, so you get really consistent, robust RF circuitry,” Carmona says.

Matched-filter baluns

One IPC offering is Johanson Technology’s line of matched-filter baluns.

A balun is an electrical device that converts between a balanced (differential) and unbalanced (single-ended) signal. The component has many forms and may include devices that also transform impedances.

Because many RF wireless chipsets have differential (two pin) outputs – an RF input and output – that connect to a single-ended antenna, the signal must be converted from differential to single-ended in a specific impedance ratio. These wireless radio frequency integrated circuits (RFICs) usually have a non-standard complex impedance that the IPCs match for optimum power efficiency. Some baluns are also combined with bandpass, low-pass, or high-pass filters.

To accomplish an impedance conjugate match, Johanson Technology works with chipset original equipment manufacturers (OEMs) to create specific baluns with matching part numbers for each chip. Reference designs during development simplify and speed up chipset adoption.

Markets

Because IPCs require much less board space, engineers can design miniaturized devices with RF circuitry and create smaller form-factor products.

Beyond size, a smaller PCB can also impact product aesthetics, allowing slim, low profiles. Eliminating components on a 10:1 or greater basis also reduces overall device weight.These factors have major implications for next-generation smart wearables (rings, bracelets, shoes, jeans, shirts, and other apparel), implantable medical devices, and portable electronics.

Johanson Technology
https://www.johansontechnology.com