Biofilm Dislodging using the High-Throughput Sonicator UIP400MTP
High-throughput sonicators designed for multi-well plates and 96-well plates are essential tools in biofilm research, particularly for dislodging and removing biofilms from wells, tubes, pins, and Petri dishes. These sonicators facilitate the processing of numerous samples simultaneously, improving efficiency and consistency. The Hielscher high-throughput sonicator UIP400MTP is the ideal for sonicating multi-well plates, 96-well plates, several tubes or Petri dishes. The uniform ultrasonic vibration and cavitation reliably removes biofilms from the bottom of the plate. This makes the multi-well plate sonicator UIP400MTP the ideal sample preparation tool for biofilm removal, biofilm assays and research of bacteria and microbes.
Multiwell-Plate Sonicator UIP400MTP for Biofilm Dislodgment and Removal in High-Throughput Sample Preparation:
- Bacterial biofilms
- Eukaryotic biofilms
- Fungal biofilms
- Archaeal biofilms
- Polymicrobial biofilms / Mixed-species biofilms
- Dental biofilms (plaque)
- Marine biofilms
- Environmental biofilms
- Medical device biofilms
- Cultured cell suspensions
UIP400MTP high-throughput sonicator for biofilm removal from multi-well plates, 96-well plates, tubes and Petri dishes.
This Hielscher sonicator is ISO certified, UL, RoHs and CE compliant. It has 24/7 operation capability for high-throughput workflows.
Biofilm Dislodging and Removal using Sonication
The Hielscher high-throughput sonicator UIP400MTP is tailored for the reliable and uniform sonication of multi-well and 96-well plates. Essential in biofilm research, the multi-well plate sonicator UIP400MTP offers a reliable technology for dislodging and removing biofilms from multi-well plates, tubes and Petri dishes. Allowing for high-throughput sample preparation, the UIP400MTP enhances the efficiency and consistency of biofilm assays and other related studies, which makes this sonicator indispensable in both academic and industrial microbiology research.
Biofilm removal using sonication: For biofilm dislodging, biofilms are allowed to form in the wells of the multi-well plate under controlled conditions. The plate is then placed in the sonicator, where parameters such as time, amplitude, and frequency are set according to the specific requirements of the biofilm type and experimental goals. The ultrasonic waves generate cavitation within each well, effectively disrupting the biofilm matrix and dislodging the cells.
Ultrasonic sample prep for biofilms: In biofilm assays, high-throughput sonicators have several applications. After sonication, dislodged biofilm cells can be collected and quantified using various assays such as crystal violet staining, viable cell counting, or biomass measurement. Additionally, biofilms can be treated with antimicrobial agents followed by sonication to assess the efficacy of these agents in disrupting biofilms. Dislodged biofilm cells are also useful for genomic and proteomic studies, as they can be used for DNA, RNA, and protein extraction for further molecular analysis.
High-throughput sonicators offer numerous advantages. They allow simultaneous processing of multiple samples, saving time and labor, and ensure uniform treatment of all wells, reducing variability and improving reproducibility of results. These devices are also suitable for large-scale studies, enabling high-throughput screening of biofilm formation, disruption, and treatment.
(A) Plate containing TSB with 2% glucose used for biofilm formation, cell recovery, and determination of MIC and MBCB; (B) Lid with pins for formation of staphylococcal biofilms.
The biofilm cells formed on the pins were dislodged using the Hielscher sonicator UIP250MTP for 5 min in 96-well plates containing fresh culture medium for recovery of the cells.
(Picture and study: ©de Oliveira et al., 2016)
96-well plate sonicator UIP400MTP for the sonication of microtiter and multiwell plates
Importance of Ultrasonic Biofilm Removal in Biofilm Research
Ultrasonic biofilm removal is crucial in biofilm research due to its effectiveness, consistency, efficiency, and versatility. It ensures thorough and uniform disruption of biofilms, facilitating accurate and reproducible analysis in various biofilm assays. These assays, including crystal violet staining, viable cell counting, ATP bioluminescence, XTT reduction, microscopic analysis, and nucleic acid/protein extraction, provide comprehensive insights into biofilm formation, viability, structure, and response to treatments.
Sonication dislodges and removes biofilm gently from solid substrates such as plate bottoms, pins, pegs, or coverslips.
- Effective Disruption of Biofilms:
Cavitation: Ultrasonic waves create cavitation bubbles that generate strong shear forces upon collapsing, effectively disrupting the complex structure of biofilms.
Thorough Dislodging: Ensures that biofilm cells are thoroughly dislodged from surfaces, which is crucial for accurate quantification and analysis. - Consistency and Reproducibility:
Uniform Treatment: High-throughput sonicators provide uniform ultrasonic energy distribution across all samples, reducing variability and improving reproducibility.
Standardization: Enables standardization of biofilm disruption protocols, leading to more reliable and comparable results. - Efficiency:
High-Throughput Capability: Allows simultaneous processing of multiple samples, saving time and increasing throughput in biofilm studies.
Automation: Can be easily integrated into automated workflows, further enhancing efficiency and reducing manual labor. - Preservation of Viability and Integrity:
Controlled Conditions: Parameters such as time and intensity can be finely tuned to disrupt biofilms without compromising cell viability or integrity, which is important for subsequent assays. - Versatility:
Broad Applicability: Suitable for various types of biofilms (bacterial, fungal, mixed-species) and compatible with different surfaces and materials used in biofilm research.
The multi-well plate sonicator UIP400MTP offers numerous benefits, which make them an indispensable tool in high-throughput sample preparation.
Hielscher Ultrasonics offers various sonicator models including probe-type sonicators, non-contact sonicators and high-throughput sonicators. Get in contact with us now! Our technical experts will be glad to discuss your processing requirements with you and recommend you the most suitable sonicator for your biofilm application.
Microtiter plate sonicator UIP400MTP is used to dislodge biofilms for research
Literature / References
- FactSheet UIP400MTP Multi-well Plate Sonicator – Non-Contact Sonicator – Hielscher Ultrasonics
- 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.
Frequently Asked Questions around Biofilm Research
What is the most common Scaffold for Biofilms?
The most common scaffold for biofilms in laboratory research is the microtiter plate. Microtiter plates, particularly the 96-well format, are widely used due to their versatility, ease of use, and compatibility with various analytical techniques.
What is the best 96-well Plate for Biofilms?
The best 96-well plate for biofilm studies depends on several factors including the type of biofilm being studied, the experimental setup, and the specific requirements of the research. However, some commonly recommended 96-well plates for biofilm studies are:
- Corning® Costar® 3596 96-well Clear Flat Bottom Plate: This plate is often used due to its high optical clarity, which is beneficial for imaging and spectrophotometric assays.
- Nunc™ MicroWell™ 96-Well Plates: These plates are known for their uniform well dimensions and high optical clarity. They are available in various surface treatments to support different types of cell culture.
- Greiner Bio-One CELLSTAR® 96-Well Microplate: This plate is frequently used for biofilm studies because of its consistent well-to-well quality and availability in different surface coatings, such as tissue culture-treated or low-binding surfaces.
- Black/Clear Bottom 96-Well Plates: For studies requiring fluorescence or luminescence measurements, black plates with clear bottoms (e.g., Corning® 3603) are ideal as they minimize cross-talk between wells and improve signal detection.
- Hydrophobic or Hydrophilic Coated Plates: Depending on the nature of the biofilm and the microorganism, plates with specific coatings that promote or inhibit adhesion can be beneficial. For instance, hydrophobic plates may be suitable for biofilms formed by certain bacteria.
- Biofilm-Specific Plates: Some manufacturers offer plates specifically designed for biofilm research. These plates often have enhanced surface properties that promote biofilm formation and are optimized for biofilm assays.
When selecting a 96-well plate for biofilm studies, consider the following criteria:
- Material and Surface Treatment: Choose a plate material and surface treatment that supports the adhesion and growth of the biofilm-forming microorganisms you are studying.
Optical Clarity: For assays involving optical measurements, ensure the plate has high optical clarity. - Well Design: Consider the shape and depth of the wells, which can affect biofilm growth and the effectiveness of subsequent treatments or measurements.
- Compatibility with Equipment: Ensure the plate is compatible with your sonicator, incubator, and any other equipment used in your study. The Hielscher multi-well plate sonicator is compatible with all standard microtiter and multi-well plates.
Ultimately, the best choice will depend on the specific requirements of your biofilm research and the characteristics of the microorganisms involved.
What are Typical Biofilm Assays used in Research?
In research and diagnostics, various types of assays are utilized to investigate and classify biofilms. Sonication is a common sample preparation step to remove the biofilm from its solid substrate making the biofilm available for assays and analysis.
- Crystal Violet Staining:
Purpose: Quantifies total biofilm biomass.
Method: Biofilms are stained with crystal violet dye, and the dye is then solubilized and measured spectrophotometrically.
Application: Used to assess biofilm formation and the effects of antimicrobial agents. - Viable Cell Counting (CFU Assay):
Purpose: Determines the number of viable biofilm cells.
Method: Dislodged biofilm cells are serially diluted and plated on agar to count colony-forming units (CFUs).
Application: Evaluates biofilm cell viability and antimicrobial efficacy. - ATP Bioluminescence:
Purpose: Measures metabolic activity of biofilm cells.
Method: ATP levels are quantified using a bioluminescent assay.
Application: Indicates biofilm viability and metabolic state. - XTT Reduction Assay:
Purpose: Assesses cellular metabolic activity.
Method: XTT reagent is reduced by metabolically active cells to form a colored formazan product, which is quantified spectrophotometrically.
Application: Used to evaluate biofilm cell viability and the effect of treatments. - Microscopic Analysis:
Purpose: Visualizes biofilm structure and composition.
Method: Techniques include light microscopy, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM).
Application: Provides detailed insights into biofilm architecture, distribution, and the effects of treatments. - DNA, RNA, and Protein Extraction:
Purpose: Analyzes genetic and proteomic profiles of biofilm cells.
Method: Biofilm cells are lysed to extract nucleic acids and proteins for downstream analysis (e.g., PCR, qPCR, sequencing, proteomics).
Application: Studies gene expression, genetic diversity, and protein expression in biofilms. - Biofilm Inhibition and Eradication Assays:
Purpose: Screens compounds for their ability to prevent biofilm formation or eradicate established biofilms.
Method: Biofilms are treated with potential anti-biofilm agents, and disruption is assessed using the aforementioned assays.
Application: Drug discovery and development of anti-biofilm therapies.
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