Isolation of PBMCs and cell populations for improved genomic analysis

Isolation of target cells increases the resolution of bulk and single-cell genomic analysis, by excluding the background signals coming from non-target cells, and thereby helps to save time and decrease sequencing costs. MACS® Technology allows the fast and easy immunomagnetic isolation of single cells from solid tissues and blood samples, helping to prevent changes in gene expression for more reliable and high resolution genomic analysis. MACS Technology also includes automated solutions to upscale, standardize, and speed up your magnetic cell separations for reproducible results.

 

Genomic analysis of magnetically isolated cells

High purity of immunomagnetic isolated PBMCs
Cellular stress responses were prevented when PBMCs were isolated directly from whole blood with immunomagnetic MicroBeads

Ficoll®-free isolation directly from whole blood yields highly pure PBMCs and prevents upregulation of cellular stress-related genes

Gentle direct isolation of PBMCs from whole blood using immunomagnetic beads prevents activation of cellular stress responses compared to density gradient centrifugation methods. In addition to that, the automated PBMC isolation procedure is standardized and reproducible, improving purity of the obtained PBMCs population.

Highly pure PBMCs directly isolated from whole blood using the Whole Blood PBMC Isolation Kit, human and Sedimentation Kit 2*

 Sample 1Sample 2Sample 3
GranulocytesWhole Blood PBMC Isolation Kit0,27%0,29%0,2%
Ficoll®4,9%2,41%1,58%
ErythrocytesWhole Blood PBMC Isolation Kit0,12%0,06%0,06%
Ficoll®2,59%37,81%21,94%

* Please contact us for product and ordering information.

Magnetic isolation of TILs increases the sensitivity of single-cell immunoprofiling
Magnetic isolation of TILs increases the sensitivity of single-cell immunoprofiling

Isolation of TILs from human tumors significantly increases the resolution of single-cell immune profiling

Magnetic isolation improves the resolution of single-cell RNA sequencing of T and B cell receptors, highlighting clonotypes otherwise poorly or altogether unrepresented in the bulk sample.

Required prodcucts

REAlease CD4/CD8 TIL MicroBead Kit, human 
gentleMACS Octo Dissociator with Heaters

Related PDFs

Isolation of TILs after tumor dissociation significantly improves the sensitivity of single-cell immune profiling (application note)
Improved RNA sequencing analysis of magnetically isolated neurons
Magnetic neuron isolation improves the resolution of single-cell analysis of different neuronal types in adult mouse brain

Isolation of neurons from adult mouse brain increases resolution of single-cell RNA sequencing analysis

Magnetic isolation of neurons after dissociation of adult mouse brain substantially increases the resolution of single-cell gene expression profiling, allowing the identification of several neuronal subpopulations which otherwise could not be distinguished.

Related PDFs

Automated mouse brain dissociation and neural cell isolation increases efficiency and sensitivity for single–cell gene expression profiling (scientific poster)
Successful detection of SNP zygosity after tumor cell isolation 

Isolation of tumor cells significantly increases the sensitivity of SNP analysis after NGS

Tumor cell isolation enables accurate detection of single nucleotide polymorphisms (SNP) in solid tumors for the analysis of loss of heterozygosity using next generation sequencing (NGS).

Required products

Tumor Cell Isolation Kit, human
autoMACS® Pro Separator

Related PDFs

Untouched tumor cell isolation allows for subsequent enrichment of rare cancer stem cells and improves NGS analysis (application note)

Enrichment of carcinoma cells from FFPE lung adenocarcinoma specimens increases the sensitivity of mutation analysis

Enrichment of carcinoma cells from FFPE lung adenocarcinoma specimens with low estimated tumor cell content, increases significantly the allele frequency of detected driver mutations compared to macrodissection.

Increased sensitivity of NGS-based mutation analysis of FFPE lung adenocarcinoma specimens by carcinoma cell enrichment.

Specimen IDPre-processing prior DNA extractionConcentration of extracted DNA (ng/ul)DNA input for NGSDriver mutation identifiedCoverageAllele frequency
Specimen 1 (70% tumor cells)Macrodissection48.840EGFR: c.2236_2250del;
p.E746_A750del		147152.3%
 Enriched tumor cells38.840EGFR: c.2236_2250del;
p.E746_A750del		252259.9%
Specimen 2 (10% tumor cells)Macrodissection4.7820KRAS: c.34G>T; p.G12C38811%
 Enriched tumor cells0.4620KRAS: c.34G>T; p.G12C62328%
Specimen 3 (< 5% tumor cells)Macrodissection5.6440KRAS: c.34G>T; p.G12C4051%
 Enriched tumor cells2.1240KRAS: c.34G>T; p.G12C9517%

Required products

FFPE Tissue Dissociation Kit
gentleMACS Octo Dissociator with Heaters

Related PDFs

Enrichment of tumor cells from FFPE lung adenocarcinoma resection specimens increases sensitivity of mutation analysis (application note)
 

How to isolate target cells using MACS Technology

Magnetic cell isolation with MACS Technology
Magnetic cell isolation with MACS Technology

Target cell isolation with MACS® Technology

MACS Technology enables the magnetic separation of cell populations by targeting surface antigens with specific antibodies that are conjugated to superparamagnetic beads. Labeled cells are magnetically retained in a separation column, from which they can easily be eluted. The fast and gentle isolation minimizes cellular changes and ensures high viability. Explore the different solutions for target cell isolation with MACS Technology here.

Related information

Principle of MACS MicroBeads 
Magnetic cell separation strategies with MACS Technology 

MACS Handbook chapters

Magnetic cell separation
Automated cell isolation using the autoMACS Pro Separator
Automated cell isolation using the autoMACS Pro Separator

Automated cell separation solutions for reproducible results

Whether working with only few samples or large sample cohorts, automation helps to streamline cell separation. The MACS Cell Separation instruments allow different levels of automation according to the sample throughputs. In particular, the autoMACS Pro Separator provides full automation for magnetic cell separations using MACS Technology, while minimizing temperature changes and user-dependent variation.

See all autoMACS Pro Separator features

Cell enrichment to decrease flow sorting time

The frequency of certain target cell populations, such as tumor infiltrating leukocytes, can be very low in the starting material, and lead to very long flow sorting times, which add up the higher thenumber of samples to be sorted, increasing the risk for gene expression changes to happen. Pre-enrichment of low abundant cell types with MACS Technology prior flow sorting allows much faster acquisition times and higher quality of the cells for genomic analysis.

Pre-enrichment of target cells reduces the flow acquisition time

Cell typeCells to analyzeEvents to collectFlow cytometry time/sample*Total flow cytometry time**
CD4+ T cells
Bulk5,0007.96×10⁶ 66.3 min>10 h
Isolated***5,0005.41×10⁴ 0.5 min ~11 min
CD8+ T cells
Bulk50002.80×10⁶ 23.3 min>3.5 h
Isolated***50004.37×10⁴ 0.4 min~10 min
T cells
Bulk10,0008.13×10⁵ 6.8 min>1 h
Isolated***10,0003.24×10⁴ 0.3 min<10 min
* Flow rate: 2,000 events/s
** Considering 9 samples (3 experimental groups × 3 replicas/group). Includes 45 s automated mixing and rinsing between samples on the MACSQuant® Instrument
*** Isolation using CD8 (TIL), CD4 (TIL), or CD4/CD8 (TIL) MicroBeads, respectively

Isolation of CD4+, CD8+, and pan T cells from different mouse tumor models using mouse CD4 (TIL) MicroBeads, CD8 (TIL) MicroBeads, and CD4/CD8 (TIL) MicroBeads dramatically decreases time of analysis.

Required products

CD4 (TIL) MicroBeads, mouse
CD8 (TIL) MicroBeads, mouse
CD4/CD8 (TIL) MicroBeads, mouse
autoMACS Pro Separator

Further information

Workflow overview

The complete workflow for cell and nuclei preparation for genomics
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video preview
Webinar

Preparation of cells and nuclei for genomic analysis
Instrument information

autoMACS Pro Separator

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