Hypersonic levitation and spinning: paving the way for enhanced single-cell analysis via contactless tissue dissociation.

Abstract

In the realm of single-cell analysis, effective tissue dissociation is a cornerstone yet often hampered by the drawbacks of traditional methods. Mechanical and enzymatic dissociation methods suffer from long processing times, reduced cell viability, and the loss of rare cell populations. Herein, we introduce a revolutionary tissue dissociation approach, Hypersonic Levitation and Spinning (HLS), which capitalizes on a uniquely designed triple-acoustic resonator probe. This probe enables the target tissue sample to levitate and execute a 'press-and-rotate' operation within a confined flow field, generating microscale 'liquid jets' that exert precise hydrodynamic forces in a non-contact manner. Through this mechanism, the shear forces on the tissue are enhanced, facilitating rapid and efficient dissociation while safeguarding cell integrity. We have further developed an automated tissue dissociation apparatus that integrates dissociation, fluid replacement, filtration, and output functions. Our comprehensive experiments on human renal cancer tissue dissociation, including flow cytometry, primary cell culture, immunofluorescence, and single-cell RNA sequencing, clearly demonstrate that Hypersonic Levitation and Spinning method not only greatly outperforms traditional techniques in tissue utilization (90% in 15 minutes vs. 70% in 60 minutes) and dissociation rate but also excels in maintaining high cell viability (92.3%) and preserving rare cell populations. This non-contact, gentle yet highly efficient dissociation method holds immense promise in diverse fields such as cell biology, single-cell sequencing, and precision medicine, expanding the scope of tissue dissociation technology and its applications.