Easy to Use Automated Magnetic Cell Sorting

August 21, 2018

Memorial University of Newfoundland
Division of BioMedical Sciences, Faculty of Medicine
Research Assistant

Overall

Performance

Ease-of-Optimization

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Company:

MIltenyi Biotec

Product Name:

autoMACS Pro Separator

Catalog Number:

130-092-545

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This review is for the autoMACS Pro, which has been discontinued and replaced by the autoMACS Neo.

The autoMACS Pro Separator from Miltenyi Biotec allows full automation of magnetic cell sorting of your desired cell population of interest for your research. It has the capability to isolate cells from your purified heterogeneous cell sample, but also directly from primary tissue sources such as whole blood or bone marrow. If you are already experienced with MACS products for manual cell separation, the transition to automated cell sorting is straight forward as several of the reagents used by both techniques are the same. MACS Microbead technology in combination with the autoMACS Pro Separator offers an alternative or complement to flow cytometry based sorting, and is compatible with several downstream applications including but not limited to cell culture, functional assays, flow cytometry, PCR, and large scale gene expression analysis. Our lab routinely used the autoMACS separator to obtain highly pure immune subsets from peripheral blood mononuclear cells.

Experimental Design and Results Summary

Application

Magnetic cell separation

Protocol Overview

Peripheral blood mononuclear cells (PBMC) were isolated by either Ficoll or using a specialized vacutainer (BD CPT tube). Positive selection of immune cell subsets was performed by suspending recovered PBMC in ~80 μl of buffe per 1 x 10^7 cells and by incubating with 22 μl antibody-labelled microbeads (Miltenyi Biotec) for 15 min at 4 °C. PBMC were washed with 10 ml of buffer and thoroughly re-suspended in 2 ml to ensure a single-cell suspension. Separation of microbead-labelled cells was done using an autoMACS Pro Separator CD19+ (B cells), CD8+ (cytotoxic T cells), CD14+ (monocytes), and CD4+ (helper T cell) subsets were isolated from the same PBMC sample in sequential order using CD19, CD8, CD14, and CD4 microbeads, respectively. Immune subset purity was determined using flow cytometric analysis.

Tips

It is important to have a single cell population of cells to avoid clogging and adequate separation. There are different programs that are designed to prioritize cell recovery or purity. Thus some preliminary experiments may be required to evaluate which automated separation protocol provided by the instrument is suitable for your experiments in order to achieve your desired balance of cell recovery and purity. Separated cell populations should be routinely checked by flow cytometry to ensure the purity of the isolated cells.

Features Summary

The autoMACS Pro Separator is a benchtop magnetic cell sorter that allows gentle sorting of more than ten million cells per second from a sample of up to 4×10⁹ total cells. Eliminate tedious manual handling steps. Save time with fully automated multisample cell separation. Automated startup, cleaning, and shutdown. Optimized washing, decontamination, and maintenance programs. Continuous monitoring of buffers and waste levels. The autoMACS Pro Separator comes with 12 pre-set separation programs for optimized and reproducible cell isolation. The appropriate program is chosen depending on the separation strategy, target cell frequency, and level of antigen expression. A high-resolution touchscreen with intuitive menus allows for straightforward and easy operation. The status menu provides continuous monitoring of sample processing. Sort cells in 3 simple steps: 1. Scan reagents using the barcode reader. 2. Position samples and reagents on instrument. 3. Select “run” and walk away. The barcode recognizes the reagents and selects the appropriate separation and wash program. Hands-free sample processing leads to safe laboratory practices, of vital importance when dealing with hazardous samples.

DOI or PMID #

26307036

Additional Notes

N/A

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Summary

The Good

User friendly interface. Easy to use and maintain. Full automation is possible, or a combination of manual and automation is also possible. Pre-set programs that allow you to customize your separation to suit your needs. Reagents come with detailed protocols and are already optimized for use with the instrument. Ability to add attachments as user needs grow.

The Bad

Reagents and buffers can get expensive depending on frequency of use.

The Bottom Line

If your lab frequently performs cell separations to obtain pure populations of cell from heterogeneous samples, then the autoMACS Pro Separator should be seriously considered. Compared to flow cytometry-based sorting applications, sample preparation is less complicated and time consuming and less technical expertise is required.

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