The Most Overlooked Challenge in Biomedical Research: Sample Preparation

Author: Dr. Josie L. Duncan, PhD

Biomedical research is, by and large, defined by its capacity to produce accurate, reliable, and repeatable results that may inform diagnostics, prognostics, and treatment planning. Without this, research will never translate into actual patient care. While many engineering practices are used to improve the robustness of these tests and analyses, proper sample preparation is a proven method for enhancing the accuracy and reliability of clinical tests and bioanalytical processes, yet it is often overlooked. When it comes to cell-based research, from benchtop to the clinic, high-purity samples enriched for the cell type of interest greatly improve the sensitivity and reliability of the results; this is often achieved by cell sorting. 1,2  Traditional cell sorting processes such as fluorescent- and magnetic-activated cell sorting (FACS and MACS, respectively), suffer from glaring limitations such as the inability to process low-volume samples, the requirement of high cell concentration,  the inaccuracy related to cell aggregation, the reduced cell viability, and the difficulty navigating debris-laden samples.3,4 These limitations often leave the user with an inconclusive or inaccurate result both after sorting and after analysis. Improving the way samples are sorted, enriched, and purified pre-analysis may significantly impact accuracy and throughput. Reliable sorting alternatives and other sophisticated approaches to sample preparation that address the limitations of these techniques, however, are seldom available.

Unlike other technologies, the CytoR1TM Platform offers a multifaceted approach to the shortcomings of FACS and MACS – sample preparation via label-free sorting for direct subsequent analysis and improving label-based sorting techniques when used upstream -- demonstrating the dynamic opportunity to incorporate CytoRecovery’s technology into any workflow.

CytoRecovery, has developed a unique approach to address sample preparation limitations for samples of low volume and/or cell concentrations while leaving the cell in its native state – a feature that is ideal for downstream analysis. This means what you put in will the same as what you get out in terms of behavior and properties. The CytoR1 Platform can sort cells of interest without a label, removing the challenges associated with clustering, bias, and cell stress from traditional sorting techniques. From a heterogeneous sample, the CytoR1 platform can enrich cells based on size, viability, and other unique biophysical differences. When performed as a step in labs’ sample preparation protocols, CytoRecovery’s technology is compatible with a multitude of processes such as genomic sequencing, toxicant and therapeutic evaluation, and subsequent culture.

CytoRecovery is partnering with the National Institutes of Health to improve sample preparation efforts at the agency through a Research Collaboration Agreement with the Center for Human Immunology (CHI). This particular effort aims to isolate viable cells low in number from whole biopsies, peripheral blood mononuclear cells (PBMCs), and bone marrow aspirations to enable more accurate immune response evaluation. Traditional cell sorting technologies may compromise the health of the sample, alter the native state of the cell, mandate higher cell counts for accuracy, or require beads and labels followed by their subsequent removal. The CytoR1 Platform is accurate even at low cell numbers, enriches for viability, and sorts without compromise from debris without labeling any cells. Together, CytoRecovery and the NIH will conduct sample prep to isolate rare cell types from clinically relevant specimens to achieve optimal samples for downstream evaluation, culture, and genomic sequencing by enriching viable cells and immune cells from PBMCs using the CytoR1 Platform.

Sample preparation is an integral part of any cell-based research, from academia to healthcare, and its impact on the end result is inarguably apparent. The CytoR1 Platform fills the gaps left by traditional cell sorting techniques, offering its users a novel way to sort cells that optimizes sample viability, nativity, and sterility for a variety of applications.

Your Cells. As They Are. Sorted.

Key Takeaways:

  • Sample preparation is directly related to the reliability and accuracy of the subsequent analysis of that sample.
  • The CytoR1 Platform optimizes sample preparation by performing label-free, high-viability sorts, even on low volume and low cell concentration samples.
  • CytoRecovery and the NIH CHI are partnering to enhance the accuracy and robustness of immune response evaluation by enriching clinical samples for cells of interest, based on viability, size and phenotype.

References

  1. Sample Preparation: A Comprehensive Guide. (n.d.). Organomation. https://www.organomation.com/sample-preparation-a-comprehensive-guide#:~:text=Why%20is%20Sample%20Preparation%20Crucial,data%20in%20any%20analytical%20workflow
  2. Ingle, R. G., Zeng, S., Jiang, H., & Fang, W.-J. (2022). Current developments of bioanalytical sample preparation techniques in pharmaceuticals. Journal of Pharmaceutical Analysis, 12(4), 517–529. https://doi.org/10.1016/j.jpha.2022.03.001
  3. Sutermaster, B.A., Darling, E.M. Considerations for high-yield, high-throughput cell enrichment: fluorescence versus magnetic sorting. Sci Rep 9, 227 (2019). https://doi.org/10.1038/s41598-018-36698-1
  4. Majka SM, Miller HL, Helm KM, Acosta AS, Childs CR, Kong R, Klemm DJ. Analysis and isolation of adipocytes by flow cytometry. Methods Enzymol. 2014;537:281-96. doi: 10.1016/B978-0-12-411619-1.00015-X. PMID: 24480352; PMCID: PMC4143162.