Your sorted cells are stressed – here are 3 reasons why you should care!

Your sorted cells are stressed – here are 3 reasons why you should care!

Accessible for all – the NX One starts at <$100K

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Are you a researcher working with electrostatic, jet-in-air, or other FACS™ sorters/cytometers, or are you trying to move away from limiting dilution, magnetic beads, and other manual cell isolation methods? If so, you need to know about sorter-induced cellular stress (SICS1). This phenomenon could be silently sabotaging your experiments – here’s why it matters and how you can stay ahead of the curve in single-cell analysis.

#1: Sorting can mess up your cells metabolically

Conventional flow cytometers are like a life-changing roller coaster ride for your cells. Whether you’re using high (70+ psi) or low (20+ psi) sample pressures, your cells are experiencing significant metabolic disruptions (Fig. 1). Energy metabolism, redox balance, and lipid biosynthesis pathways are all affected[1-3]. Think about it – would you trust data from cells that have just been through the cellular equivalent of a theme park thrill ride?
Figure 1. Electrostatic sorting induces significant changes to the cellular metabolic profile (Modified from Choudhury et al., 2024).

#2: You might be unknowingly misinterpreting disease states

Here’s a scary thought: what if the gene expression changes you’re seeing aren’t from your experimental conditions but from the sorting process itself? SICS can alter gene expression profiles, potentially leading to misinterpretation of cellular disease states. This could mean the difference between a breakthrough and a dead end in fields like oncology or immunology. Do you really want to take that risk and potentially sacrifice all your painstaking preparation?

Figure 2. Representative images of pluripotency marker (TRA-1-60) staining on NX One sorted iPS colonies and parental iPS cell line. Nuclei were stained using Hoechst. Scale bar: 100µm.[4]

#3: Your stem cells’ potential is at stake

The risks are even higher for those working with stem cells, particularly induced pluripotent stem cells (iPSCs).  These cells’ long-term health and differentiation potential are crucial for applications like regenerative medicine. Traditional sorting methods could compromise (or inadvertently alter) the very properties that make these cells valuable. Are you willing to gamble with the future of your precious stem cells?

Figure 3. (A) Representative images demonstrating colony formation from a single iPS cell sorted by the NX One system. The inset and arrow at Day 0 show cell quantification. Transient cytosolic marker CytoTell UltraGreen was used to facilitate gating/sorting and imaging of single cells. Cells remain pluripotent based on their morphology after colony formation. (B) The colony formation rate from single iPSCs sorted by the NX One was comparable to the colony formation from manual limiting dilution control.

The solution: Embrace the microfluidic revolution

Don’t panic! There’s a game-changing solution available for every lab. Nodexus Inc.’s NX One benchtop cell sorting and live cell dispensing system is powered by microfluidic technology and is rewriting the traditional rules of cell sorting. With sample pressures of 0.7 psi (which can be 100x lower than conventional systems!), this innovative sorter offers:

  • Gentle and efficient sorting
  • Industry-leading cell viability
  • Maintained pluripotent status
  • Uncompromised differentiation potential

Accessible for all – the NX One starts at <$100K

By adopting this cutting-edge technology, you’re not just sorting cells – you’re preserving their native transcriptomic profiles and boosting their survival rates for downstream applications – all at a fraction of typical cell sorters’ cost and training requirements.

Ready to take your single-cell workflows to the next level? Don’t let SICS hold you back. Explore Nodexus’s microfluidic sorting solutions and stay at the forefront of life science research. Your cells (and your data) will thank you.

References
  1. Richardson GM, Lannigan J, Macara IG. Does FACS perturb gene expression? Cytometry A. 2015 Feb;87(2):166-75.
  2. Choudhury FK, Premkumar V, Zecha J, Boyd J, Gaynor AS, Guo Z, Martin T, Cimbro R, Allman EL, Hess S. Multiomics Characterization of a Less Invasive Microfluidic-Based Cell Sorting Technique. J Proteome Res. 2024 Feb 28.
  3. Box A, DeLay M, Tighe S, Chittur SV, Bergeron A, Cochran M, Lopez P, Meyer EM, Saluk A, Thornton S, Brundage K. Evaluating the Effects of Cell Sorting on Gene Expression. J Biomol Tech. 2020 Jun 22:jbt.20-3103-004.
  4. Neuronal patterning of NX One sorted pluripotent monoclonal iPSCs – https://nodexus.com/wp-content/uploads/2024/03/NX-One-Neuronal-Patterning-of-Monoclonal-iPSCs.pdf

Explore the NX One

Compact design for operation on the benchtop or inside the biosafety cabinet.

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Laser and rugged optics for particle sizing and fluorescence measurements
Wireless software for monitoring runs and visualizing data
Microfluidic cartridge for gentle, contamination-free sorting and dispensing. See it in action
Front-facing fluidics compartment for internal storage and easy access
Dispensing of sorted cells into microplates or collection tubes
Dispensing of sorted cells into microplates or collection tubes
Laser and rugged optics for particle sizing and fluorescence measurements
Wireless software for monitoring runs and visualizing data
Microfluidic cartridge for gentle, contamination-free sorting for cell sorting and dispensing. See it in action
Front-facing fluidics compartment for internal storage and easy access