Preparation of single-cell and single-nuclei suspensions for genomic analysis

The generation of single-cell or single-nuclei suspensions from solid tissues is the most critical step prior to single-cell or single-nuclei sequencing. gentleMACS Technology represents a gentle, yet efficient approach to either extract viable cells or nuclei from different solid tissues. Whether tissues are fresh or frozen, extraction of nuclei is fast and easy with the Nuclei Extraction Buffer. Protocols for tissue dissociation and nuclei extraction have been optimized for speed and efficiency to give reliable results in downstream genomic analyses.

Single-cell analysis of dissociated whole mouse brain
Distinct cell population identification after neonatal mouse brain dissociation

Single-cell analysis of dissociated whole mouse brain

Clear distinction of different neural cell populations from neonatal mouse brain in single-cell gene expression analysis, including all different major neural populations.

Analysis of tumor heterogeneity
Cell types identified after tumor dissociation

Analysis of tumor heterogeneity

Single-cell gene expression analysis of mouse tumors enables the clear identification of the different cellular populations present in the tissue. 

Preparation of single-cell suspensions from solid tissue

Single-cell suspensions from fresh tissues can be easily generated using gentleMACS Technology, which combines gentle mechanical disruption with mild enzymatic digestion. gentleMACS Technology allows retrieval of different cell types present in the tissue, using automated and standardized protocols for reproducible results. 

 

Video protocols for tissue dissociation

Find out how to standardize tissue dissociation using gentleMACS™ Technology step-by-step.

Preparation of single-nuclei suspensions from solid tissue

Single-nuclei suspensions can be easily generated from different types of solid tissues using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociators. This combination allows a gentle mechanical disruption of the tissues and an efficient cell lysis to release intact nuclei from cells in only seven minutes and of up to eight samples in parallel at low temperature.

Preparation of single nuclei suspensions from mouse brain and tumors
Preparation of single nuclei suspensions from mouse brain and tumors

High nuclei yields from mouse brain and tumors 

Single-nuclei suspensions obtained from fresh and snap frozen mouse brain and syngeneic tumors show high numbers of extracted nuclei per mg of dissociated tissue.

Preparation of single nuclei suspensions from snap frozen mouse tissue and OCT embedded human tumors
Preparation of single nuclei suspensions from snap frozen mouse tissue and OCT embedded human tumors

High nuclei yields from different snap frozen mouse tissues and OCT embedded human tumors 

Single-nuclei suspensions obtained from different snap frozen mouse tissues and OCT embedded human tumors show high numbers of extracted nuclei per mg of dissociated tissue.

Single nuclei suspension from mouse brain
Single nuclei suspension from mouse brain

Microscopy analysis of nuclei extracted from mouse brain 

Intact nuclei from snap-frozen mouse brain extracted using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator with Heaters.

Video
gentleMACS Dissociators: Reliability built in

Preparing single-cell and single-nuclei suspensions is fast and efficient with gentleMACS Dissociators. Genomic analysis results are more dependable because of the standardization and automation of gentleMACS Technology protocols.

Further information