Ultrasonic Disintegration of Cells
Ultrasonication is an effective means of disintegrating cell structures. Therefore, sonicators are widely used in laboratories to break open cells, extract intracellular molecules, proteins and organelles for research and analysis. On industrial scale, ultrasonic disintegration and lysis is used to isolate molecules from cell factories or to promote the digestion of biomass.
What is Ultrasonic Disintegration?
Ultrasonic disintegration, also known as ultrasonic homogenization, is a process that uses high-intensity, low-frequency ultrasound waves to break down cell walls and disrupt molecular structures in a liquid medium. This technique is commonly used in various scientific and industrial applications for several purposes:
Interruzione delle cellule: Ultrasonic disintegration is widely used in cell biology and molecular biology to disrupt cell membranes, releasing cellular contents such as proteins, nucleic acids, and organelles. This is useful for extracting intracellular components for analysis or for lysing cells in microbiology and biotechnology processes.
- Omogeneizzazione: It helps in the uniform mixing of components in a sample, especially when dealing with immiscible liquids or when trying to achieve a consistent blend of materials.
- Protein Extraction: In biology, proteomics life science, the analysis of proteins is a very common task. Before proteins can be analysed in assays, they must be extracted from the cell interior and isolated. Sonicators are the most widely used method for protein extraction.
- Frammentazione del DNA: DNA and RNA are distinct types of nucleic acids that store and encode genetic information in cells. When DNA and RNA are analyzed, the long strands must sometimes be fragmented, a process that can be reliably and efficiently done by sonication.
- Sample Preparation: In research and analysis, sample preparation is a common procedure before various analytical techniques. Ultrasonic disintegration can help to dissolve or disperse samples, which may improve the accuracy and reproducibility of analyses.
Advantages of Ultrasonic Disintegration
Why using a probe-type sonicator for disintegration, cell disruption and the extraction of intracellular molecules and proteins? A sonicator or ultrasonic dismembrator offers numerous advantages that makes sonication the superior technology when compared to other disintegration methods such as high-pressure homogenization, ball milling or microfluidization.
- Non-Thermal: Ultrasonic disintegration is a non-thermal method, which means it does not rely on heat to break down materials. This is advantageous for applications where high temperatures could degrade heat-sensitive samples.
- Precise and Controlled: The process can be controlled with high precision, allowing for specific disruption, mixing, or particle size reduction.
- Rapid and Efficient: Ultrasonication is generally a fast and efficient method, making it suitable for high-throughput applications.
- Reduced Chemical Usage: In many cases, ultrasonic disintegration can reduce the need for harsh chemicals or organic solvents, which can be environmentally friendly and reduce the risk of chemical contamination.
- No Milling Media, No Nozzles: Alternative disintegration techniques, such as ball/bead milling or high-pressure homogenizers, come with disadvantages. Ball/bead milling necessitates the use of milling media (beads or pearls), which must be laboriously separated and cleaned. High-pressure homogenizers have nozzles that are prone to clogging. In contrast, ultrasonic homogenizers are easy to use, highly reliable and robust, requiring very little maintenance.
- Versatilità: It can be applied to a wide range of materials, including bacteria, plant cells, mammalian tissue, algae, fungi etc. making it a versatile technique in various fields.
Scalabilità: The ultrasonic technique can be scaled up for industrial processes, making it suitable for both laboratory and large-scale production applications.
The Working Principle of Ultrasonic Disintegration and Cell Disruption
Ultrasonication generates alternating high-pressure and low-pressure waves in the exposed liquid. During the low-pressure cycle, the ultrasonic waves create small vacuum bubbles in the liquid that collapse violently during a high-pressure cycle. This phenomenon is termed cavitation. The implosion of the cavitation bubble causes strong hydrodynamic shear-forces that cause first sonoporation and subsequent the efficient disruption of cell structures. Intracellular molecules and organelles are completely released into the solvent.
Disintegrazione ultrasonica delle strutture cellulari
The shear forces can disintegrate fibrous, cellulosic material into fine particles and break the walls of the cell structure. This releases more of the intra-cellular material, such as starch or sugar into the liquid. In addition to that the cell wall material is being broken into small debris.
This effect can be used for fermentation, digestion and other conversion processes of organic matter. After milling and grinding, ultrasonication makes more of the intra-cellular material e.g. starch as well as the cell wall debris available to the enzymes that convert starch into sugars. It does also increase the surface area exposed to the enzymes during liquefaction or saccharification. This does typically increase the speed and yield of yeast fermentation and other conversion processes, e.g. to boost the ethanol production from biomass.
Use Ultrasonic Disintegration – Reliably and Efficiently at Any Scale
Hielscher sonicators are available with different power ratings and processing capacities. Whether you want to sonicate small biological samples from a few microlitres to a few litres or need to process large cell or biomass streams for production, Hielscher Ultrasonics will offer you the most suitable ultrasonic dismembrator for your biological application.
- lab scale for 1mL to approx. 5L e.g. UP400St with 22mm sonotrode
- bench top scale at approx. 0.1 to 20L/min e.g. UIP1000hdT with 34mm sonotrode and flowcell
- production scale starting at 20L/min e.g. UIP4000hdt o UIP16000hdT
La tabella seguente fornisce un'indicazione della capacità di trattamento approssimativa dei nostri ultrasuoni da laboratorio:
Dispositivi raccomandati | Volume di batch | Portata |
---|---|---|
UIP400MTP Sonicatore per piastre a 96 pozzetti | piastre multipozzetto / microtiter | n.a. |
Corno a tazza a ultrasuoni | CupHorn per fiale o becher | n.a. |
GDmini2 | reattore a microflusso a ultrasuoni | n.a. |
VialTweeter | 0,5-1,5 mL | n.a. |
UP100H | 1 - 500mL | 10 - 200mL/min |
UP200Ht, UP200St | Da 10 a 1000 ml | Da 20 a 200 mL/min |
UP400St | 10 - 2000mL | 20 - 400mL/min |
agitatore a ultrasuoni | n.a. | n.a. |
Please use the form below, if you would like to receive more information regarding the use of ultrasonic devices for the purpose of the disintegration of cells. We will be glad to assist you.
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The table below gives you an indication of the approximate processing capacity of our industrial ultrasonicators:
Volume di batch | Portata | Dispositivi raccomandati |
---|---|---|
200mL to 5L | 0.05 to 1L/min | UIP500hdT |
1 to 10L | 0Da .1 a 2L/min | UIP1000hdT |
5 to 20L | 0,2 - 4L/min | UIP2000hdT |
10 - 100L | 2 - 10L/min | UIP4000hdt |
Da 15 a 150L | Da 3 a 15L/min | UIP6000hdT | n.a. | 10 - 100L/min | UIP16000 |
n.a. | più grande | cluster di UIP16000 |
Letteratura / Riferimenti
- Nico Böhmer, Andreas Dautel, Thomas Eisele, Lutz Fischer (2012): Recombinant expression, purification and characterisation of the native glutamate racemase from Lactobacillus plantarum NC8. Protein Expr Purif. 2013 Mar;88(1):54-60.
- Brandy Verhalen, Stefan Ernst, Michael Börsch, Stephan Wilkens (2012): Dynamic Ligand-induced Conformational Rearrangements in P-glycoprotein as Probed by Fluorescence Resonance Energy Transfer Spectroscopy. J Biol Chem. 2012 Jan 6;287(2): 1112-27.
- Claudia Lindemann, Nataliya Lupilova, Alexandra Müller, Bettina Warscheid, Helmut E. Meyer, Katja Kuhlmann, Martin Eisenacher, Lars I. Leichert (2013): Redox Proteomics Uncovers Peroxynitrite-Sensitive Proteins that Help Escherichia coli to Overcome Nitrosative Stress. J Biol Chem. 2013 Jul 5; 288(27): 19698–19714.
- Elahe Motevaseli, Mahdieh Shirzad, Seyed Mohammad Akrami, Azam-Sadat Mousavi, Akbar Mirsalehian, Mohammad Hossein Modarressi (2013): Normal and tumour cervical cells respond differently to vaginal lactobacilli, independent of pH and lactate. ed Microbiol. 2013 Jul; 62(Pt 7):1065-1072.