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Sample preparation

1 Sample preparation

Successful experiments strongly depend on the quality of the starting material, for example, samples of dissociated tissue. Effective and reproducible preparation of tissue samples can be challenging when using manual methods. For this reason, Miltenyi Biotec has developed the gentleMACS™ Portfolio that enables gentle, yet effective automated tissue dissociation or thorough homogenization with reproducible results.

gentleMACS Dissociator Technology combines mechanical tissue dissociation using gentleMACS Dissociators and gentleMACS Tubes with enzymatic digestion based on dedicated Tissue Dissociation Kits. All components of this technology work in unison to yield the maximal amount of viable single cells while preserving cell functions and characteristics, including surface epitopes.
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gentleMACS™ Dissociators: Start smart!


Get your research off to the right start. Rely on excellent sample preparation with the gentleMACS Dissociator family. Start your experiment with standardized and reproducible tissue dissociation with the gentleMACS Dissociator, the gentleMACS Octo Dissociator, or the gentleMACS Octo Dissociator with Heaters for fully automated tissue dissociation. 

MACS Handbook:

Spleen (mouse)

Lymph node (mouse)

Tumor tissue (mouse)

Tumor tissue (human)

Brain (mouse)

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1.1 Mechanical disruption: gentleMACS Dissociators and Tubes

gentleMACS Dissociators enable fast and easy tissue dissociation and homogenization with minimal hands-on time. At the heart of gentleMACS Dissociator Technology are the gentleMACS Tubes. They come in two versions specifically designed for the generation of single-cell suspensions or cell homogenates from tissue.
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1.1.1 gentleMACS Dissociators
Three types of gentleMACS Dissociators are available, supporting different needs.
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The gentleMACS Dissociator family includes gentleMACS Octo Dissociator, gentleMACS Dissociator, and gentleMACS Octo Dissociator with Heaters (left to right).

The gentleMACS Dissociator allows for automated dissociation or homogenization of two tissue samples in parallel. More than 30 pre-defined programs are available for processing of the most frequently used tissues, including tumor, neonatal brain, neonatal heart, lung, spleen, and intestine.

The gentleMACS Octo Dissociator enables automated processing of eight samples in parallel or sequentially. Different tissue types can be handled simultaneously and new samples can be added at any time. In addition to utilizing pre-defined programs, users can customize programs to achieve optimal preparation of their specific samples.

The gentleMACS Octo Dissociator with Heaters offers all the advantages of the gentleMACS Octo Dissociator. Additionally, the Heaters facilitate enzymatic digestion directly on the instrument for fully automated tissue dissociation.
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1.1.2 gentleMACS Tubes

gentleMACS Tubes have been designed to ensure effective dissociation and homogenization of tissue samples.

Tissue dissociation

The purple-cap C Tube has been developed specifically for the preparation of viable single-cell suspensions used in cell separation, cell culture, and cell analysis experiments.

 

Tissue homogenization

The orange-cap M Tube is designed for thorough sample homogenization enabling subsequent molecular and microbiology analysis.
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Function and benefits of gentleMACS C Tubes in detail.

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1.2 Enzymatic digestion: Tissue Dissociation Kits

MACS Tissue Dissociation Kits and gentleMACS Dissociators and Tubes are perfectly geared to each other. The combination of gentle mechanical and enzymatic tissue dissociation preserves all cell characteristics and functions for reliable and meaningful downstream analysis. Pre-defined enzyme solutions and lot-to-lot consistency of enzyme activities ensure reproducible and user-independent results.

Miltenyi Biotec researchers tested whether the proteases included in the Tumor Dissociation Kits for human and mouse tissue have an effect on the integrity of cell surface proteins. To this end, the presence of more than 200 epitopes was analyzed by flow cytometry after incubation of cells with the enzymes. The vast majority of the cell surface epitopes tested was well preserved during enzymatic treatment.
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1.3 Tissue storage

Oftentimes tissue samples cannot be processed immediately when they become available. To enable flexible timing of tissue dissociation without compromising cell yield or viability, Miltenyi Biotec developed the Tissue Storage Solution. This solution enables storage of fresh organ and tissue samples for up to 48 hours. During this time, cell viability is not affected and no undesired effects like cell activation or apoptosis induction have been observed. Tissue Storage Solution was tested and validated on a variety of human and mouse tissues including tumors, skin, heart, spleen, brain, and skeletal muscle.
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Comparison of tissues stored in MACS Tissue Storage Solution or competitor products. Numbers indicate the cell count per gram of tissue. 

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