Ultrasonic Cell Detachment
Ultrasonic cell detachment is a highly effective and reliable sample preparation technique used in various fields of cell biology and biotechnology. Using ultrasonic waves, the multi-well plate sonicator UIP400MTP detaches adherent cells from culture surfaces and prepares cell suspensions for downstream applications. Sonication offers several advantages over traditional enzymatic, chemical and mechanical detachment techniques, making it a valuable tool for researchers.
Principle of Ultrasonic Cell Detachment
Ultrasonic cell detachment relies on the use of power ultrasound to disrupt the interactions between cells and the substrate they are attached to. Ultrasonic waves generate microbubbles, acoustic cavitation and vibrations in the culture medium, producing mechanical forces that gently but effectively dislodge the cells. This process is typically carried out using the non-contact sonicator UIP400MTP for multi-well plates.
Non-contact sonicator UIP400MTP for cell detachment
Detaching cells is crucial when culturing adherent cells. While trypsinization is the most commonly used technique, it can decrease cell viability by damaging the cell membrane and extracellular matrix. To improve cell culture efficiency, it is important to minimize this damage. Ultrasonic cell detachment is a highly effective and reliable enzyme-free technique. This makes the UIP400MTP microplate sonicator an excellent alternative for enzyme-based cell detachment. Sonication applies acoustic cavitation and agitation in a serum-free medium, which gently dislodges the cells without harming them.
Comparison of Ultrasonic Cell Detachment with Traditional Techniques
| Advantages | Ultrasonic Detachment | Enzymatic Detachment | Chemical Detachment |
|---|---|---|---|
| Cell Viability | High | Medium | Variable |
| Speed | Rapid (minutes) | Moderate (minutes to hours) | Variable (minutes to hours) |
| Preservation of Surface Markers | Excellent | Can be altered | Can be altered |
| Scalability | High | Medium | Variable |
| Residue-Free | Yes | No (enzyme residue) | Variable (chemical residue) |
| Equipment Cost | One-time investment, no proprietary disposables, no re-occuring costs | Re-occuring costs | Re-occuring costs |
| Ease of Optimization | Easy | Easy | Variable |
The multi-well plate sonicator UIP400MTP offers numerous benefits for high-throughput sample preparation such as cell detachment.
This Hielscher sonicator is ISO certified, UL, RoHs and CE compliant. It has 24/7 operation capability for high-throughput workflows.
Below, you find an exemplary instruction for cell detachment using the multi-well plate sonicator UIP400MTP.
Equipment and Materials for Ultrasonic Cell Detachment
- Non-Contact Sonicator for Multi-well Plates UIP400MTP: This high-throughput sonicator is the key tool. The plate sonicator UIP400MTP is suitable for any standard multi-well and microtiter plates as well as for Petri dishes. Ultrasound waves provide the necessary mechanical energy for cell detachment.
- Cell Culture Plates or Petri Dish: Standard vessels used to grow adherent cell cultures.
- Culture Medium: The liquid medium in which cells are grown and maintained.
- Sterile PBS (Phosphate-Buffered Saline): Used for washing cells before detachment.
- Collection Tubes: For collecting the detached cell suspension.
Protocol for Ultrasonic Detachment using the Plate Sonicator UIP400MTP
- Preparation
– Ensure all equipment is clean and sterile to prevent contamination.
– Pre-warm the culture medium and PBS to the appropriate temperature (usually 37°C). - Cell Washing
– Aspirate the culture medium from the cell culture flask or plate.
– Gently wash the cells with sterile PBS to remove any residual medium and detached cells. - Sonication
– Add a small volume of PBS or fresh culture medium to the flask or plate to cover the cell monolayer.
– Place the Petri dish or plate into the UIP400MTP sonicator.
– Sonicate for a specified duration, typically ranging from a few seconds to a few minutes, depending on the cell type and the strength of attachment. - Collection of Cell Suspension
– After sonication, observe the cells under a microscope to ensure detachment.
– Gently pipette the cell suspension to collect the detached cells.
– Transfer the suspension to a collection tube. - Post-Detachment Processing
– Centrifuge the cell suspension if necessary to concentrate the cells.
– Resuspend the cells in fresh culture medium or buffer for downstream applications.
Notes: The parameters (such as sonication time and intensity) need to be optimized for different cell types to avoid cell damage. Some delicate cell types may be more sensitive to ultrasonic treatment, requiring careful optimization.
Practical Applications of Ultrasonic Cell Detachment
Ultrasonic cell detachment is used in various research and clinical settings:
- Flow Cytometry: Preparing single-cell suspensions for analysis.
- Cell Counting and Viability Assays: Detaching cells for accurate counting and viability assessments.
- Subculturing: Transferring cells from one culture vessel to another.
- Molecular Biology Experiments: Isolating cells for DNA, RNA, and protein extraction.
Why Should I Replace Cell Scraping by Cell Detachment with the UIP400MTP Microplate Sonciator?
Replacing cell scraping with cell detachment using the UIP400MTP microplate sonicator offers researchers several advantages. Unlike manual scraping, the precisely controllable sonication with the UIP400MTP ensures consistent and gentle cell detachment, minimizing mechanical damage and preserving cell viability. The UIP400MTP provides precise control over sonication parameters, enabling uniform treatment across all wells and eliminating variability between samples. This method is faster, more efficient, and scalable, making it ideal for high-throughput applications while maintaining reproducibility and the integrity of sensitive cell cultures. Find the comparative study here!
96-well plate sonicator UIP400MTP for the sonication of microtiter and multiwell plates
96-well plate sonicator UIP400MTP: The ultrasound-transmitting liquid surround all wells and provide uniform sonication of each sample
Common Cell Types for Ultrasonic Cell Detachment
- Fibroblasts:
Commonly used in tissue engineering and wound healing research.
Adherent cells that require detachment for passaging and experiments. - Epithelial Cells:
Used in studies of tissue barriers, cancer research, and drug testing.
Require detachment for analysis and subculturing. - Endothelial Cells:
Important for vascular research and studies on angiogenesis.
Adherent cells that need detachment for in vitro assays. - Stem Cells:
Including mesenchymal stem cells and induced pluripotent stem cells.
Require gentle detachment methods to maintain viability and pluripotency. - Cancer Cell Lines:
Widely used in cancer research and drug development.
Adherent cells that require regular detachment for experimental manipulation. - Hepatocytes:
Used in liver function studies and drug metabolism research.
Require detachment for in vitro models and assays. - Keratinocytes:
Predominant cell type in the epidermis and commonly used in skin research, wound healing studies, and cosmetic testing.
Like other adherent cells, keratinocytes grow attached to the surface of culture flasks or plates and must be detached for various experimental procedures, including passaging, analysis, and subculturing. - Macrophages:
Often require detachment when cultured in vitro.
Macrophages are adherent cells, which typically attach to the surface of culture flasks or plates. To collect them for downstream applications such as analysis, subculturing, or experiments, they need to be detached from the culture surface.
Literature? References
- FactSheet UIP400MTP Multi-well Plate Sonicator – Non-Contact Sonicator – Hielscher Ultrasonics
- Lauren E. Cruchley-Fuge, Martin R. Jones, Ossama Edbali, Gavin R. Lloyd, Ralf J. M. Weber, Andrew D. Southam, Mark R. Viant (2024): Automated extraction of adherent cell lines from 24-well and 96-well plates for multi-omics analysis using the Hielscher UIP400MTP sonicator and Beckman Coulter i7 liquid handling workstation. Metabomeeting 2024, University of Liverpool, 26-28th November 2024.
- UIP400MTP-Multi-well-Plate-Sonicator-Infographic
- De Oliveira A, Cataneli Pereira V, Pinheiro L, Moraes Riboli DF, Benini Martins K, Ribeiro de Souza da Cunha MDL (2016): Antimicrobial Resistance Profile of Planktonic and Biofilm Cells of Staphylococcus aureus and Coagulase-Negative Staphylococci. International Journal of Molecular Sciences 17(9):1423; 2016.
- Martins KB, Ferreira AM, Pereira VC, Pinheiro L, Oliveira A, Cunha MLRS (2019): In vitro Effects of Antimicrobial Agents on Planktonic and Biofilm Forms of Staphylococcus saprophyticus Isolated From Patients With Urinary Tract Infections. Frontiers in Microbiology 2019.
Facts Worth Knowing
What is Cell Detachment?
Cell detachment is the process of separating adherent cells from the surface of their culture vessel, enabling their collection and preparation for further experimental procedures or analysis. This can be achieved through enzymatic, chemical, mechanical, or ultrasonic methods.
What is Cell Dissociation?
Cell dissociation is the process of breaking down cell aggregates or tissues into individual, viable cells. This is achieved by disrupting the extracellular matrix and cell-cell adhesions using enzymatic, mechanical (e.g. sonication), or chemical methods. Cell dissociation is essential for various applications, including primary cell culture, single-cell analysis, and the preparation of cell suspensions for experiments and therapeutic uses.
What is the Difference between an Adherent Cell Culture and a Biofilm?
Adherent cell culture refers to the growth of cells, typically eukaryotic, that attach to a substrate in a controlled laboratory environment, relying on nutrients supplied through the culture medium. In contrast, a biofilm is a complex, naturally occurring microbial community that adheres to a surface and is encased in an extracellular matrix, with cells demonstrating collective behavior, chemical gradients, and enhanced resistance to external stresses. While adherent cell cultures are artificial and used for research purposes, biofilms are dynamic ecosystems found in natural or engineered environments.
Sonication with the UIP400MTP is a highly efficient, enzyme-free technique to detach adherent cells and to dislodge biofilms. Read more about the advantages of biofilm dislodgment from microplates and Petri dishes using the UIP400MTP multi-well plate sonicator!
What are the Advantages of Ultrasonic Cell Detachment?
- Gentle on Cells: Ultrasonic detachment is less harsh compared to enzymatic methods (like trypsinization) and mechanical scraping, preserving cell viability and surface markers.
- Rapid and Efficient: The process is quick, often taking only a few minutes, and can efficiently detach cells even from large culture surfaces.
- Scalability: Suitable for both small-scale laboratory applications and larger biotechnological processes.
- No Enzyme Residue: Eliminates the need for enzymes, reducing the risk of altering cell surface proteins or introducing contaminants.