Enhancing Routine Cell Analysis & Sorting with the Nodexus NX One PRO and Slingshot Biosciences’ TruCytes™ Cell Mimics
In the rigorous cell analysis and sorting world, researchers are continuously challenged to streamline workflows, improve accuracy, and validate methodologies to achieve reliable results. Optimizing these processes has never been more critical with the field pushing the boundaries of what’s possible. Emerging technologies are reshaping how scientists approach cell sorting, introducing tools that enhance efficiency, precision, and direct visualization.
A recent collaboration combines Nodexus’ cutting-edge cell sorting technology with Slingshot Biosciences’ innovative cell mimics. This partnership highlights an ultra-simple and highly reproducible approach to boost benchtop cell analysis and sorting capabilities and validate workflows, providing researchers with robust tools to meet the growing demands of cell analysis.
The Power of Precision: Redefining Cell Sorting with the NX One PRO
The NX One PRO is a state-of-the-art microfluidic cell sorter, expertly designed to provide researchers with a seamless blend of precision and gentleness in cell sorting applications. Equipped with up to three lasers, this easy-to-use system delivers exceptional accuracy in distinguishing and sorting cells while drastically minimizing the physical stress typically associated with traditional sorting methods. The NX One PRO’s gentle microfluidic approach with <1 PSI pressure enhances sorting efficiency. It allows researchers to retain the native characteristics of cells, making it an ideal tool for accessible cell analysis in fields ranging from immunology and oncology to developmental biology.
Cell Mimics: Reliable Controls for Cell Therapy
The Slingshot Biosciences’ TruCytes™ CD34 kit includes non-biohazardous cell mimics that replicate critical biological samples such as cord blood and bone marrow aspirate. These CD34 and CD45-expressing controls are ideal for research fields like hematology, regenerative medicine, and immunotherapy. Slingshot’s CD34 cell mimics are also representative of pre- and post-CD34 enrichment samples with two populations of 90% (CD34 high) and 1.5% (CD34 low) expressing mimics. They enable precise standardization in cell therapy manufacturing and streamline workflows by providing relevant controls to save precious biological samples for drug development, while enhancing quality control and assay validation processes.
Slingshot’s ViaComp® Flex viability cell mimics are a convenient and reliable control to measure cell health. These mimics bind to DNA intercalating dyes (such as 7-AAD and PI) or amine-reactive dyes (such as Zombie Green or Live/Dead Fixable dyes) to simulate the staining of cells. These engineered mimics offer several advantages over traditional biological controls, such as consistent size and properties, stable antibody binding, long-term storage capability, and batch-to-batch reliability.
Synthetic Biology Meets Benchtop Cell Sorting
In a series of experiments designed to demonstrate the power of cell mimics and benchtop cell analysis and sorting as part of a quality-controlled rare cell isolation process, we first combined TruCytes™ CD34 cell mimics (10-12 µm) with ViaComp® Flex beads (20 µm) to evaluate the NX One PRO’s ability to discriminate between control particles based solely on their size (Figure 1).
Figure 1. (A) Brightfield image of TruCytes™ and ViaComp® Flex beads. (B) Distinct Forward Scatter- Height peaks were observed for TruCytes™ and ViaComp® Flex beads, highlighting the NX One PRO’s ability to distinguish between the sizes. Panel 1B presents forward scatter data (FSC-H), which enables alignment-, calibration-, and label-free discrimination between the two bead populations.
The smaller TruCytes™ population appears as a separate peak from the larger ViaComp® Flex beads. This distinct separation in forward scatter signal showcases the NX One PRO’s high-resolution capabilities in size-based discrimination, proving effective for applications requiring precise sorting based on cell or particle size. The NX One PRO offers software-controlled gain adjustment for samples requiring even higher resolution to discriminate between populations. In the next experiment, the NX One PRO demonstrated high precision and accuracy in sorting rare cell mimic populations, as shown in Figure 2.
Figure 2. TruCytes™ cell mimics decorated with CD34 and CD45 were stained with CD34 PE-conjugated antibody (Panel 2A) and CD45 FITC-conjugated antibody (Panel 2B). In Panel 2C, the forward scatter height (FSC-H) and fluorescence (FL2) analysis indicate effective separation between the stained and unstained populations, with unstained comprising 99% of the samples and only 1% being CD34-PE+. Conventional “gating” or selection of populations using polygonal/freeform gates can also be seen in Panel 2C. These gates may be saved and re-imported for future experimental runs to minimize end-user setup and readily enable automation of these control workflows. Panel 2D highlights the deposition accuracy for CD34 high (90% target) and low (~1.5% target) abundance samples.
This figure underscores the NX One PRO’s high sensitivity and capability in discriminating, and ultimately isolating, rare cell populations based on scatter and fluorescence. The demonstrated results of ~90% deposition accuracy confirm the NX One PRO’s ability to achieve consistent, precise sorting for either high-frequency or rare cell populations of interest, further supporting its suitability for applications requiring reliable cell sorting.
Implications for Your Research
This collaborative study validates the NX One PRO’s advanced capabilities in high-purity sorting of rare cell populations and precise size-based discrimination. By pairing accessible tools like the NX One family of benchtop sorters (which include single- and multi-laser capabilities and now, with the NX One MAX, the ability to sort organoids, spheroids, cardiomyocytes, adipocytes, and more up to 200 μm in diameter) with cell mimics from Slingshot Biosciences (TruCytes™ CD34 Low/High and ViaComp® Flex), researchers can utilize consistent, biohazard-free controls that simplify the optimization and standardization of cell sorting protocols. These innovations address critical challenges in cell analysis, providing a versatile platform that enhances workflow efficiency, standardizes processes, and enables accessible, high-fidelity sorting for numerous applications, from basic research to therapeutic development, at an accessible price point, compact footprint, and ease-of-use level fit for every cell biology laboratory.
