Single Nuclei Isolation
Single Nuclei Isolation - Related Videos
09:09
Single Nuclei Isolation from Coronary Endarterectomy Tissue of Coronary Artery Bypass Graft Patients
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2026
Coronary endarterectomy tissue samples were processed to isolate single nuclei using an optimized dissociation protocol. The integrity of the isolated nuclei was validated through three complementary approaches: trypan blue exclusion staining to assess membrane integrity, DAPI-based confocal microscopy to confirm nuclear morphology, and flow cytometry to quantify nuclear yield and purity.
05:59
Isolation of Nuclei from Human Intermuscular Adipose Tissue and Downstream Single-Nuclei RNA Sequencing
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Cited by 2 •
2024
The biology of intermuscular adipose tissue (IMAT) is largely unexplored due to the limited accessibility of human tissue. Here, we present a detailed protocol for nuclei isolation and library preparation of frozen human IMAT for single nuclei RNA sequencing to identify the cellular composition of this unique adipose depot.
10:12
Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue
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Cited by 61 •
2012
Cardiac nuclei are isolated via density sedimentation and immunolabeled with antibodies against pericentriolar material 1 (PCM-1) to identify and sort cardiomyocyte nuclei by flow cytometry.
07:03
Isolation of Adipose Tissue Nuclei for Single-Cell Genomic Applications
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Cited by 12 •
2020
This publication describes a protocol for the isolation of nuclei from mature adipocytes, purification by fluorescence-activated sorting, and single-cell level transcriptomics.
06:38
Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing
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Cited by 69 •
2018
Here, we present a protocol to rapidly isolate high-quality nuclei from the fresh or frozen tissue for downstream massively parallel RNA sequencing. We include detergent-mechanical and hypotonic-mechanical tissue disruption and cell lysis options, both of which can be used for isolation of nuclei.
FAQs
Mechanical disruption is intended to break the plasma membrane while leaving the nucleus intact. This distinction allows nuclear contents to be recovered without requiring complete preservation of the original cell. The released nuclei can then enter a stabilizing buffer, creating a suspension suitable for downstream genetic and molecular analyses rather than losing fragile cellular material during whole-cell dissociation.
These steps improve the quality of the nuclear suspension in different ways. Stabilizing buffer receives the nuclei after release, filtration removes larger debris, and density-based centrifugation can enrich the nuclear fraction. Together, they reduce unwanted material in the preparation and produce a more suitable input for molecular profiling, including analyses of DNA, chromatin, or RNA.
The approach reduces dependence on fresh tissue and can be applied to frozen or archived material, according to the overview. It is also useful when tissue structure or cell fragility makes whole-cell dissociation problematic. By analyzing nuclei rather than requiring intact cells, the method can reduce effects associated with dissociation and preserve access to cell-type-resolved genetic information.
An isolated nuclear suspension can support several molecular readouts, including DNA, chromatin, and RNA profiling. These measurements can be used for single-nucleus sequencing and for examining gene expression, epigenetic states, and genomic variation. In genetics, the major value is the ability to connect these features with individual cellular or nuclear populations rather than interpreting the tissue only as a bulk sample.
A typical workflow begins with mechanical disruption of the tissue or cells to break the plasma membrane. The released nuclei are transferred into a stabilizing buffer, passed through a filter to remove debris, and may then undergo density-based centrifugation for enrichment. The resulting suspension becomes the preparation for downstream DNA, chromatin, RNA, or single-nucleus sequencing analyses.
The core components described are the starting tissue or cells, a mechanical disruption step, stabilizing buffer, a filter, and equipment for density-based centrifugation when enrichment is needed. Their sequence matters: nuclei must be released before filtration and possible density separation. The resulting preparation is intended to limit debris and provide a usable input for molecular and genetic assays.
It is particularly valuable when researchers need cell-type-resolved information from frozen, archived, or structurally complex samples, or when fragile cells are poorly suited to whole-cell dissociation. Single-nucleus analyses can reveal differences in gene expression, epigenetic states, and genomic variation across nuclear populations, extending genetic profiling to samples that are difficult to process as intact cells.