Replace Cell Scraping with the UIP400MTP High-throughput Sonicator
The detachment and extraction of adherent cell lines from multi-well plates for multi-omics analysis is a daily task in labs. High-throughput cell detachment using the multi-well plate sonicator UIP400MTP replaces manual cell scraping leading to higher yields of RNA, total lipids and total polar metabolites. A new method integrates the Hielscher UIP400MTP sonicator with the Beckman Coulter i7 liquid handling workstation, enabling high-throughput, reproducible, and efficient processing of cells for RNA, metabolite, and lipid extraction. The presented method excels traditional manual cell scraping methods by achieving superior reproducibility and yield across various cell types and experimental conditions.
Streamline Cell Detachment with the Microplate Sonicator UIP400MTP
Adherent cell culture systems play a critical role in toxicological and biomedical research. Within this context, Cruchley-Fuge et al. (2024) addressed a significant challenge in the PrecisionTox project focused on leveraging omics technologies for chemical hazard evaluation. The project aimed for a high-throughput analysis of thousands of samples treated with diverse chemicals. To meet this demand, the researchers developed an automated workflow combining the UIP400MTP sonicator with established biphasic extraction protocols for liquid chromatography-mass spectrometry (LC-MS) analysis. Their study evaluates the efficacy of the UIP400MTP Multi-Well Plate Sonicator in detaching adherent cells compared to manual scraping and other traditional methods.
Detaching adherent cell cultures from any standard microplate – at high-throughput with the UIP400MTP sonicator
Streamlining Multi-Omics: Automated Adherent Cell Extraction with the UIP400MTP
The researcher team of Laura Cruchley-Fuge from the University of Birmingham utilized three human cell lines: HepG2 (liver cancer cells), HepaRG (differentiated hepatocyte-like cells), and H295R (adrenal cancer cells). These cells were cultured in 24-well and 96-well plates and exposed to test chemicals such as aflatoxin B1 and forskolin.
Experimental Design:
- Phase 1: Optimization of the UIP400MTP sonicator’s power settings and comparison with manual cell scraping and sonic water baths. HepG2 cells were employed to evaluate RNA, metabolite, and lipid recovery.
- Phase 2: Integration of the UIP400MTP into a biphasic extraction workflow using the Beckman Coulter i7 system. Validation was conducted using HepaRG and H295R cells.
Extraction Workflow: The workflow encompassed chemical exposures in multi-well plates, cell detachment using the UIP400MTP, and biphasic extraction via the Bligh & Dyer (B&D) method. LC-MS analysis was performed using a Thermo Scientific Orbitrap Exploris 120 for lipophilic and polar compounds. The B&D method, a gold standard for lipid quantification, involves a two-step extraction with methanol, chloroform, and water, followed by lipid quantification in the chloroform phase.
UIP400MTP microplate sonicator facilitates the detachment of adherent cell lines from multi-well plates and Petri dishes
Results:
- Phase 1: Optimal sonication conditions were identified at 60% power.
The UIP400MTP yielded the highest RNA recovery with exceptional reproducibility compared to manual scraping and sonic baths.
Polar metabolite recovery was consistent across methods, while lipid recovery was notably superior with the UIP400MTP. - Phase 2: Validation on HepaRG and H295R cells demonstrated high reproducibility in lipidomics and metabolomics data, as indicated by tightly clustered PCA scores.
Aflatoxin B1 and forskolin treatments were effectively distinguished from controls, underscoring the method’s sensitivity and reliability.
Microplate Sonicator UIP400MTP for high-throughput cell detachment
“The Hielscher UIP400MTP sonication device provides a high-quality and reproducible alternative approach to the “gold standard” of manual cell scraping, leading to higher yields of RNA, total lipids and total polar metabolites.” (Cruchley-Fuge et al., 2024)
Cruchley-Fuge et al. highlight the advantages of the UIP400MTP sonicator for adherent cell processing. By replacing manual scraping, this method enhances reproducibility, throughput, and yield, making it an invaluable tool for large-scale studies like PrecisionTox. The UIP400MTP’s integration into automated workflows not only reduces variability but also streamlines labor-intensive processes, enabling high-quality multi-omics data acquisition.
The work of Cruchley-Fuge et al. (2024) facilitates and streamlines the processing of adherent cell cultures for multi-omics analysis. The integration of the UIP400MTP sonicator with automated workflows ensures consistent and efficient sample preparation, making it ideally suited for high-throughput toxicological research.
The UIP400MTP is not an ultrasonic bath. It'a a high intensity cup horn for focused sonication. This powerful non-contact sonicator delivers a uniform sonication across all wells of your standard well-plate. You have precise control over amplitude, power, and pulsing. A built-in timer and temperature probe ensures consistent results. The UIP400MTP plate sonicator cools samples with water bath (external chiller optional).
Design, Manufacturing and Consulting – Quality Made in Germany
Hielscher ultrasonicators are well-known for their highest quality and design standards. Robustness and easy operation allow the smooth integration of our ultrasonicators into industrial facilities. Rough conditions and demanding environments are easily handled by Hielscher ultrasonicators.
Hielscher Ultrasonics is an ISO certified company and put special emphasis on high-performance sonicators featuring state-of-the-art technology and user-friendliness. Of course, Hielscher ultrasonicators are CE compliant and meet the requirements of UL, CSA and RoHs.
High-throughput cell detachment without cell scraping – the UIP400MTP sonicator facilitates lab work.
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.
- Dreyer J., Ricci G., van den Berg J., Bhardwaj V., Funk J., Armstrong C., van Batenburg V., Sine C., VanInsberghe M.A., Marsman R., Mandemaker I.K., di Sanzo S., Costantini J., Manzo S.G., Biran A., Burny C., Völker-Albert M., Groth A., Spencer S.L., van Oudenaarden A., Mattiroli F. (2024): Acute multi-level response to defective de novo chromatin assembly in S-phase. Molecular Cell 2024.
- Mochizuki, Chika; Taketomi, Yoshitaka; Irie, Atsushi; Kano, Kuniyuki; Nagasaki, Yuki; Miki, Yoshimi; Ono, Takashi; Nishito, Yasumasa; Nakajima, Takahiro; Tomabechi, Yuri; Hanada, Kazuharu; Shirouzu, Mikako; Watanabe, Takashi; Hata, Kousuke; Izumi, Yoshihiro; Bamba, Takeshi; Chun, Jerold; Kudo, Kai; Kotani, Ai; Murakami, Makoto (2024): Secreted phospholipase PLA2G12A-driven lysophospholipid signaling via lipolytic modification of extracellular vesicles facilitates pathogenic Th17 differentiation. BioRxiv 2024.
- Cosenza-Contreras M, Seredynska A, Vogele D, Pinter N, Brombacher E, Cueto RF, Dinh TJ, Bernhard P, Rogg M, Liu J, Willems P, Stael S, Huesgen PF, Kuehn EW, Kreutz C, Schell C, Schilling O. (2024): TermineR: Extracting information on endogenous proteolytic processing from shotgun proteomics data. Proteomics. 2024.
- 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.
Frequently Asked Questions
What is Cell Detachment?
Cell detachment in research refers to the process of separating adherent cells from the surface of a culture vessel or substrate. This is typically done to harvest cells for downstream applications such as analysis, subculturing, or cryopreservation. Detachment can be achieved using enzymatic methods (e.g., trypsin), chemical agents (e.g., EDTA), mechanical methods (e.g., scraping), or physical techniques like sonication, depending on the cell type and research requirements.
How do you Detach Adherent Cells?
Detaching adherent cells using sonication involves the application of focused ultrasound waves to disrupt the cell-surface adhesion within a controlled environment. Specifically, the UIP400MTP microplate sonicator achieves this by generating localized mechanical vibrations that break the bonds between cells and the culture surface. Key steps include:
- Preparation: Cells are grown in multi-well plates and may be exposed to specific chemicals as part of the experimental design.
- Sonication: The sonicator UIP400MTP is programmed with optimized settings (e.g., 60% power) to ensure effective detachment without damaging the cells or compromising biomolecule integrity.
- Temperature Control: The device maintains temperature stability to prevent heat-induced cell or molecular degradation during the process.
- Post-Detachment: Detached cells are subjected to downstream extraction protocols, such as the Bligh & Dyer biphasic method, for the recovery of RNA, lipids, and metabolites.
This method is superior to manual scraping due to its automation, reproducibility, and ability to process high-throughput samples efficiently.
What is Non-Damaging Cell Detachment?
Non-damaging cell detachment refers to the process of separating adherent cells from their substrate without compromising cell viability, integrity, or functionality. It is achieved using gentle methods such as controlled sonication or enzyme-free solutions.
Avoiding cell destruction is critical to preserving the cells’ structural and molecular characteristics, which are essential for accurate downstream applications like multi-omics analysis, functional assays, or therapeutic use. Damaged cells can release intracellular contents, potentially confounding experimental results or compromising sample quality.
What is the Advantage of Enzyme-Free Cell Detachment?
Enzyme-free cell detachment offers several advantages, including preserving cell surface proteins and receptors, maintaining cell viability, and avoiding potential enzymatic damage to biomolecules. This approach is particularly beneficial for sensitive downstream applications, such as flow cytometry, proteomics, or functional assays, where enzymatic alterations could compromise data quality or experimental outcomes. Additionally, enzyme-free methods are often more reproducible and can be adapted for high-throughput workflows.
Hielscher Ultrasonics manufactures high-performance ultrasonic homogenizers from lab to industrial size.