FixNCut: Single-cell genomics through reversible tissue fixation and dissociation

Author:

Jiménez-Gracia LauraORCID,Marchese DomenicaORCID,Nieto Juan C.ORCID,Caratù GinevraORCID,Melón-Ardanaz ElisaORCID,Gudiño VictoriaORCID,Roth Sara,Wise Kellie,Ryan Natalie K,Jensen Kirk B.,Hernando-Momblona Xavier,Bernardes Joana P.,Tran Florian,Sievers Laura Katharina,Schreiber Stefan,van den Berge Maarten,Kole Tessa,van der Velde Petra L.,Nawijn Martijn C.,Rosenstiel Philip,Batlle EduardORCID,Butler Lisa M.ORCID,Parish Ian A.ORCID,Plummer JasmineORCID,Gut IvoORCID,Salas AzucenaORCID,Heyn HolgerORCID,Martelotto Luciano G.ORCID

Abstract

AbstractThe use of single-cell technologies for clinical applications requires disconnecting sampling from downstream processing steps. Early sample preservation can further increase robustness and reproducibility by avoiding artifacts introduced during specimen handling. We present FixNCut, a methodology for the reversible fixation of tissue followed by dissociation that overcomes current limitations. We applied FixNCut to human and mouse tissues to demonstrate the preservation of RNA integrity, sequencing library complexity, and cellular composition, while diminishing stress-related artifacts. Besides single-cell RNA sequencing, FixNCut is compatible with multiple single-cell and spatial technologies, making it a versatile tool for robust and flexible study designs.

Publisher

Cold Spring Harbor Laboratory

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