Capture and Labeling of cancer cells using Aria

This application note describes the use of the new Fluigent Aria – a software-assisted instrument capable of delivering up to 10 different solutions – for the automation of a complete protocol of capture and labeling of MDA-MB-231 breast cancer cells including surface treatment, injection of antibodies-coated beads, and cell suspension, and immunostaining steps.

We have demonstrated the use of Aria and its related software for the automated delivery of different liquids for the capture and labeling of breast cancer cells using a complex microfluidic setup. We also demonstrate how Aria can adapt to specific protocols by making use of its specific software features, allowing for the optimization of protocols.

Context

Cell capture and labelling are usually time consuming processes that can last for several hours, and often require exposure to multiple fluids. As an example, typical staining protocols usually require more than 5 consecutive solutions. Cell labelling, cell manipulation, drug screening or perfusion also demand very precise injection volumes with the use of controlled flow rates (ranging from 1 μL/min to 1 000 μL/min) in order to perform reproducible experiments. Smooth fluid control is also recommended as uncontrolled flow rates can create high shear stresses, subsequently affecting cell functional properties and damaging samples. Aria is the perfect instrument to address these concerns and automate fluid delivery protocols.

Challenge

We describe the automation of a rare cell capture and detection method including surface treatment, beads and cells solution injections, and immunostaining using precise volume control using Aria and its related automation software. We use the EPHESIA microfluidic chip from Institut Curie and MDA-MB-231 breast cancer cell line as a demonstration of system function and utility.

webinar fluid delivery fluigent

Conclusion

We have demonstrated the use of Aria and its related software for the automated delivery of different liquids for the capture and labelling of breast cancer cells using a complex microfluidic setup. This protocol included 10 different liquid injections, surface treatment, beads injection, cells injection, capture and labelling in a sequential and automated manner. We also demonstrate how Aria can adapt to specific protocols by making use of its specific software features, allowing for the optimization of complex protocols.

Aria provides key features for performing this application as it brings to the user:

  • »  Reduced manipulation time and timed exposure to antibodies, fluorophores, DNA probes with automated and timed protocols
  • »  Reduced handling for minimal contamination< or changes tocell conditions
  • »  Higher reproducibility compared to manual methods, hencemore reliable results

References

  1. Autebert, J. et al. 2015 High purity microfluidic sorting and analysis of circulating tumor cells: towards routine mutation detection. Lab Chip 15, 2090–2101 (2015).
  2. Kim, M. Y. et al. Tumor Self-Seeding by Circulating Cancer Cells. Cell 139, 1315–1326 (2009).
  3. Cabel, L. et al. Clinical potential of circulating tumour DNA in patients receiving anticancer immunotherapy. Nat. Rev. Clin. Oncol. 15, 639–650 (2018).
  4. Bidard, F. C. et al. Clinical validity of circulating tumour cells in patients with me- tastatic breast cancer: A pooled analysis of individual patient data. Lancet Oncol. 15, 406–414 (2014).
  5. Saias, L., Autebert, J., Malaquin, L. & Viovy, J.-L. 2011 Design, modeling and characte- rization of microfluidic architectures for high flow rate, small footprint microfluidic systems. Lab Chip 11, 822–32 (2011).
  6. Bernacka-wojcik, I. 2014 Design and development of a microfluidic platform for use with colorimetric gold nanoprobe assays. (Universidade Nova de Lisboa, 2014)

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