Novel instrumentation enables gentle and consistent cell washing for researchers across the life sciences.

This article was first published on Charles River Laboratories Eureka Blog. Please support our partners at Charles River.

“The future is now” certainly rings true for biotechnology and medicine. Gene sequencing and editing have become routine, cell biology is understood enough to turn cells into drugs, and vaccines combating a global pandemic are manufactured and delivered in less than a year. Curiously, preparing cell samples for these advances in biology rely on a technology first developed in the 19th century—the centrifuge. Fast-forward to today, my company, Curiox Biosystems has developed Laminar Wash™ technology, a next-generation alternative to the ancient cell-spinner.

The centrifuge plays such an important role in the life science laboratory because the quality of cell analysis data (e.g., flow cytometry, mass cytometry, and single-cell ‘omics) depends on extensive cell washing. The adage “garbage in, garbage out” is as popular with cell biologists as it is with data scientists. Proper cell washing leads to increased cellular resolution, as eliminating all the debris and unbound antibody, in turn yields accurate target cell identification. Unfortunately, when it comes to the centrifuge, “proper” means subjecting precious cells to multiple bouts of high gravitational forces at which astronauts would wince, altering cellular morphology and metabolism, clumping cells and shredding others apart, thus generating additional debris that require removal.

Less spinning, more cleaning

Instead of pelleting cells into such close proximity, Laminar Wash technology gently removes debris, dead cells, and excess reagent with—as the name suggests—laminar flow. This is accomplished by combining a novel “wall-less” 96-well plate design with a dual-nozzle fluidics instrument that initiates the laminar flow across each well. Laminar flow velocity is highest above the cells, removing unwanted floating waste, while negligible at the bottom, leaving cells practically undisturbed. The user indicates the number of wash cycles, and the cells are continuously cleaned, allowing the researcher to walk away. Although at two minutes per cycle, multitasking probably does not include a lunch break.

Charles River is using Laminar Wash to help characterize tumor model systems for therapy developers. Dr. Christoph Eberle, PhD, the principal scientist doing these experiments, is most interested in tumor-infiltrating lymphocytes (TILs), the immune cell combatants (or all too often, idle spectators) within the tumor microenvironment (TME), and he compares analyzing such samples as “panning for gold” whereas viable TIL cells represent the gold.

In a recent webinar , Dr. Eberle pointed out that “when comparing cell pelleting by centrifugation and manual breaking up of the cell pellet for staining, the Laminar Wash overall gives a cleaner sample.” In describing flow cytometry readout from a B16F10 tumor, “notoriously one of the messiest, and therefore hardest TIL sample to work with,” he concluded that the Laminar Wash method yielded better retention of viable cells compared to the manual processing, which help to better define immune cell populations down the gating hierarchy. Dr. Eberle has clearly found a modern tool for his gold prospecting endeavors.

Towards standardizing flow cytometry

In addition to disrupting cells, the centrifuge typically fails to deliver on another 21st century trend— laboratory automation . Automating repetitive operations both redeploys skillful researchers to less mundane tasks and represents a weapon against what Dr. Eberle calls the “reproducibility crisis,” a complication ubiquitous to the life sciences. Laminar Wash systems can be easily integrated into existing robotics solutions or can themselves complete flow cytometry workflows end-to-end. Not only did Laminar Wash improve data quality in Dr. Eberle’s lab, “it also overall simplified the workflow and saved processing time required for flow cytometric analysis.”

Laminar Wash-driven improvements in data quality and workflow efficiency are being adopted broadly across the life sciences, including front-end ing single-cell ‘omics techniques, enabling high-color flow cytometry assays , and supporting COVID-19 research (currently unpublished). Since the instruments can be operated in a biosafety cabinet and do not generate aerosols, they are ideal for researchers looking to limit exposure to infectious agents. Next-generation techniques and therapies deserve a modern solution to sample preparation. Laminar Wash technology offers a gentle, consistent, and safe alternative to the centrifuge for preparing cherished cells.

By Geoffrey Feld, Ph.D.

Marketing & Content Manager, Curiox Biosystems