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

Area of Science:

• Biotechnology
• Molecular Biology
• Genomics

Background:

• Clinical applications of single-cell technologies necessitate decoupling sample collection from downstream processing.
• Early sample preservation is crucial for enhancing robustness and reproducibility by mitigating specimen handling artifacts.

Purpose of the Study:

• To introduce FixNCut, a novel methodology for reversible tissue fixation followed by dissociation.
• To overcome current limitations in sample preservation for single-cell analyses.

Main Methods:

• Developed a reversible fixation and dissociation technique named FixNCut.
• Applied FixNCut to human and mouse tissues.
• Evaluated RNA integrity, sequencing library complexity, and cellular composition preservation.
• Assessed the reduction of stress-related artifacts.

Main Results:

• FixNCut successfully preserved RNA integrity and sequencing library complexity in both human and mouse tissues.
• The method maintained cellular composition, minimizing stress-related artifacts.
• Demonstrated compatibility of FixNCut with single-cell RNA sequencing and other single-cell and spatial technologies.

Conclusions:

• FixNCut offers a versatile solution for robust and flexible study designs in single-cell and spatial analyses.
• The methodology enhances the reliability of clinical applications by improving sample preservation and reducing artifacts.