As researchers endeavor to map the architecture of the brain, many rely on tissue dissociation approaches to prepare single-cell suspensions for downstream analysis. However, such conventional methods can involve enzymes and heat, which are known to induce transcriptional and proteotype changes in brain cells.
Choosing a more gentle approach can help researchers to effectively characterize brain cell populations while protecting the characteristics of precious tissue samples.
This whitepaper highlights a tissue dissociation technology that uses acoustic energy to generate single-cell suspensions that maintain the characteristics of important cellular subsets.
Download this whitepaper to discover:
- The importance of preserving cell surface characteristics in brain tissue samples
- An enzyme- and heat-free approach that can generate more individual cells than conventional methods
- How the latest technology can help researchers identify rare cell subpopulations
As researchers continue to map the architecture of the brain and identify rare subsets of cells with essential functions, single cell analyses of brain tissue are becoming increasingly important. Many researchers rely on tissue dissociation to prepare single cell suspensions for downstream multiomic applications (1). The conventional approach for tissue dissociation involves treatment with enzymes and sometimes heat, a process which Mattei et al. have demonstrated introduces significant transcriptional and proteotype bias in bran cells (2). Having previously demonstrated enhanced CD8+ T cell dissociation from B16 melanoma tumors collected from C57BL/6 mice, we* wanted to compare our SimpleFlow™ Bulk Lateral Ultrasound (BLU) technology to conventional enzyme and heat-based dissociation of brain tissue. BLU implements gentle dissociation with acoustic energy while maintaining samples at constant temperatures of 6°C, a fast and automated process which enables tissue dissociation into single cell suspensions in as little as 3 minutes. To assess whether our BLU single cell dissociation technology could elucidate these differences both, two independent experiments were performed at Grand Valley State University through our academic collaboration with Dr. Kristin Renkema’s laboratory. In these experiments where B16 melanoma tumors had been allowed to grow for 12-16 days, brains were collected post-harvest to yield three biological replicates per condition (N = 3) in the first experiment (E1) and five biological replicates per condition (N = 5) in a second follow-up experiment (E2). Brain tissue was dissociated using BLU or a commercially available brain tissue dissociation kit according to the manufacturer’s protocol. Single cell suspensions were then assessed by flow cytometry for several cell surface markers and for cell counts using CountBright™ absolute counting beads. Marker expression was determined by FlowJo and two-sample t-tests were performed in GraphPad Prism to determine significance of differences in means between conditions.
