Cell Isolation: Fundamental Insights

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Cell Isolation techniques are methods to separate and to transfer certain cells from a complex mixture of cells to obtain single cells or to sort the cells according to a property of choice and thus to generate a homogenous cell population. Single cell analysis is crucial across various research fields as it allows for detailed examination of individual cells, cell clusters, or subcellular compartments. This approach provides insights into cellular heterogeneity, interactions, and functions, which is essential for advancing our understanding of disease mechanisms, therapeutic potentials, and biological processes. Cell isolation enables more precise and personalized approaches in medicine and enhances research in diverse areas such as cancer, neurobiology, stem cell research, crop science, and forensics.

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Cell Isolation: Which Techniques are used?

Before starting a single cell analysis, scientists must identify and isolate single cells. There are several very different cell isolation methods available depending on the starting material, and varying on throughput as well as on sample requirements as well as downstream technologies. The starting material can be tissue, cell suspensions and cell culture.

1. Fluorescence Activated Cell Sorting (FACS): Efficient for high-throughput cell isolation based on specific markers, but it has several disadvantages. Sample preparation can be labour-intensive and requires cells to be in a single cell suspension, which may be challenging for certain tissues. The rapid flow and charge used in FACS can damage cell viability, leading to potential cross-contamination and failure in downstream analysis. Additionally, data interpretation can be complex, as accurate results depend on proper gating strategies and compensation for spectral overlap between markers. There are also limitations in detecting very rare cell populations, particularly if their fluorescence signals are weak compared to more abundant populations, which can complicate cell isolation efforts.

2. Magnetic Activated Cell Sorting (MACS): Similar to FACS, MACS is also efficient for high-throughput cell isolation but has some disadvantages. It requires a large starting number of cells and can potentially cause damage or stress to cells. Additionally, the magnetic fields used in MACS can harm some cell populations, impacting their viability and function.

3. Microfluidics: The most commonly used method in microfluidics for cell isolation is cell-affinity chromatography, which can process large numbers of cells simultaneously, significantly increasing throughput. However, shear forces and other mechanical forces within microfluidic channels can affect cell viability and function.

4. Manual Cell Picking/Micromanipulation: The technique enables the controlled separation of selected living cells from suspension, but also from fixed cells. Although it is time-consuming, it offers significant advantages for precise cell isolation. Cell picking provides a method for targeted cell isolation, while CellEctor stands out as the most flexible and gentle approach available.

5. Laser Capture Microdissection (LCM): A relatively rapid and precise method for isolating and removing specific cells from complex tissues for subsequent analysis of their RNA, DNA, protein, or metabolite content. LCM is considered the gold standard for cell isolation, especially due to its gentle, flexible, and contamination-free isolation.

1. FACS: In this process, suspended labelled cells are directed through a stream of droplets, with each droplet containing a single cell in front of a laser. The fluorescence detection system identifies the fluorescent and light scatter properties of each cell. Based on these characteristics, a charge is applied to the droplets containing cells of interest. An electrostatic deflection system then directs the charged droplets into separate collecting tubes.

2. MACS: This method involves labeling target cells with magnetic beads conjugated to specific antibodies. The labeled cells are then separated from the rest of the cell suspension using an external magnetic field.

3. Cell Picking: Target cells are observed under a microscope, and single cells are isolated using a glass capillary. This method allows for direct cell isolation by manually selecting and transferring cells into a new tube for further analysis.

4. Microfluidics: Prior to conducting experiments, cells are dissociated and then flowed into a microfluidic chip. This setup enables the separation of individual cells into different tubes, each containing a single cell.

5. LCM: This precise and gentle technique employs a laser to melt the membrane containing the target cells. An isolation cap then captures the sample without direct contact, ensuring no contamination. This method is the least provided and yields the best results for downstream analysis, highlighting its effectiveness in cell isolation.

What is the Gold Standard for Cell Isolation?

LCM is the powerful technique used to isolate single cells or a targeted cell group with extreme microscopic precision. This technology can selectively isolate single cells from various origins, such as from tissue sections, cytospins or smears, and from cell culture to perform single cell omics. LCM focuses on using a low-pulse ultraviolet laser on membrane-mounted sections of tissue. The membrane offers good optical quality without interfering with staining and the downstream analysis.

What makes LCM the gold standard for cell isolation? The combination of the MMI CellCut LCM system with the ZEISS Axio Observer inverted microscope allows for the precise and gentle isolation of single cells without compromising the spatial information of the cell’s micro environment.

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Cell Isolation System: CellCut with the ZEISS Axio Observer

What are the Benefits of LCM for Cell Isolation?

1. The most gentle isolation
LCM with the MMI CellCut and ZEISS Axio Observer features the cleanest laser on the market for dissection. The specialized laser creates an exceptionally precise cutting edge as thin as 0.3 µm, protecting the surrounding area and preserving biological information. Additionally, the sample stage moves, keeping the laser beam consistently in focus and maintaining a minimal cutting edge. This makes the MMI CellCut the most delicate method for cell isolation.

2. Check your efficiency with 100% success rate
The unique CapLift technology ensures nearly a 100% success rate in cell isolation. This technology uses an Isolation Cap with an adhesive surface that captures the sample after dissection, allowing for visual inspection of the excised sample.

3. Contamination-Free Isolation
During the cutting process, cell isolation is achieved without direct contact between the isolation cap and the sample. This is done using a sandwich technique, where the sample is placed between a membrane slide and a glass slide, ensuring the process remains entirely contamination-free.

4. High-Throughput Automation
The LCM system from MMI and ZEISS can be enhanced with an automated, high-throughput guided workflow for cell isolation. The innovative CellRobot system enables the dissection of samples in a 96-well plate in less than 10 minutes.

5. Cut it all
The cell isolation system is versatile to dissect almost any type of sample. It can handle fresh frozen tissue, FFPE material, plant tissue, living cells, smears, cytospins, and more. It can also cut through thick tissues such as teeth, bones, or forensic tapes. Even living cells can be isolated without removing the culture medium, preserving cell integrity for subsequent recultivation, cloning, or single cell analysis. The integration of a climate chamber with full environmental control is also possible.

6. Become an expert in 10 minutes
The system is incredibly user-friendly, allowing you to become a professional in just 10 minutes. The intuitive software provides a user-friendly interface, enabling detailed documentation of every cell isolation step (including images and videos).

7. Unmatched Flexibility
The LCM CellCut system is the most flexible option for cell isolation. It supports all standard objectives and imaging modes. Additionally, the system can be upgraded with a whole slide scanner, a cell picker, and AI for detecting specific cells.

8. Exceptional Imaging Quality with the ZEISS Axio Observer
Paired with the ZEISS Axio Observer, this system delivers exceptional performance with flexibility to support future growth. The ZEISS Axio Observer provides exceptional optical performance, superior resolution, and advanced imaging capabilities, ensuring that even the most delicate samples are captured with extraordinary detail.

Our Customer Success Stories for Cell Isolation

Our experience for cell isolation with MMI CellCut has been overwhelmingly positive. Its flexibility and user-friendly interface allowed seamless integration of diverse imaging and proteomic data, enhancing our analysis of tumor and neural heterogeneity.

The MMI team’s flexibility in providing ongoing support and improvements ensured a highly satisfactory user experience. CellCut achieves a 100% success rate in single cell and subcellular collection, helping us discover new immunosuppressive cell subtypes in pancreatic cancer, with significant publications forthcoming.

We highly recommend MMI for clients with single cell or subcellular applications requiring high collection success rates and minimal cellular damage, we highly recommend MMI CellCut.

Ethan Hao

CEO of BayOmics

We appreciate the resistant MMI product quality, the professional consulting, and the competent and quick service. MMI instruments are an important basis for our in-situ analysis of cellular tissue. The MMI CellCut laser microdissection for cell isolation followed by gene expression analysis is complementary to further routine methods like conventional optical microscopy (fluorescence), in-situ hybridization, and immunohistochemistry.

Danny D. Jonigk
Director of the Institute of Pathology
RWTH Aachen, Germany

Related Products

ZEISS Axio Observer
Inverted microscope that combines excellence for life science research

CellCut
LCM system for the best cell isolation experience

CellEctor
Manual and automated single cell picking

Downloads

Brochure
Benefits of LCM for Cell isolation

Specifications
Specs of CellCut LCM system

How Does MMI and ZEISS Support Researchers and Laboratories?

MMI and Zeiss collaborate to provide comprehensive support for researchers and laboratories focusing on cell isolation. Our integrated solution combines the Zeiss Axio Observer microscope with MMI’s LCM system, designed to streamline cell isolation processes. Our expert team ensures that the entire system is professionally installed at your facility, enabling seamless operation right from the start. The innovative Switchbox solution allows the ZEISS microscope to be used for various applications beyond cell isolation. This dual-use capability offers unparalleled flexibility, adapting to different research needs.

With our complete system, you receive not only cutting-edge technology but also exceptional long-term support. MMI and Zeiss are committed to providing global, high-quality service to ensure your success and satisfaction with our cell isolation solutions.

How can we Support your Cell Isolation Project?

Contact us for further questions

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Head Office Germany

Molecular Machines and Industries GmbH

Breslauer Strasse 2

D – 85386 Eching

info@molecular-machines.com

+49 89 319 048 40