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Ultrasonic cell disruption, also known as sonication, is a technique used in the laboratory to break down cell membranes. This is typically achieved through the use of an ultrasonic processor that generates high frequency sound waves. These sound waves create microbubbles in the liquid medium surrounding the cells, and the rapid growth and collapse of these bubbles results in shear forces that disrupt cell membranes and release intracellular contents. This technique is valuable in biotechnology and related fields for extracting cellular components, including proteins, DNA, and other biomolecules for further analysis or use.
Here's a simplified explanation of the process:
A sample containing cells is placed in a container (such as a test tube or beaker).
The container is then placed in an ultrasonic processor. The processor contains a vibrating probe, also known as a sonicator tip, which generates high-frequency sound waves.
These sound waves pass through the sample, creating microbubbles in the liquid medium that the cells are in.
The microbubbles rapidly expand and collapse, a process known as cavitation. The forces generated by this process disrupt the cell membranes.
The disruption of the cell membranes allows the intracellular contents to be released into the surrounding medium. These contents can then be separated, purified, and studied.
It's important to note that sonication can generate heat, which can be harmful to some cell components. Therefore, care must be taken during sonication to prevent overheating, such as using an ice bath to cool the sample or sonicating in short pulses to allow the sample to cool between rounds of sonication.
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• Cell disrupter (extraction of plant substances, disinfecting, enzyme deactivation)
• Therapeutic ultrasound, i.e. induction of thermolysis in tissues (cancer treatment)
• Decrease of reaction time and/or increase of yield
• Use of less forcing conditions e.g. lower reaction temperature
• Possible switching of reaction pathway
• Use of less or avoidance of phase transfer catalysts
• Degassing forces reactions with gaseous products
• Use of crude or technical reagents
• Activation of metals and solids
• Reduction of any induction period
• Enhancement of the reactivity of reagents or catalysts
• Generation of useful reactive species
The process of ultrasonic disruption involves several key steps:
Generation of Ultrasonic Waves: Ultrasonic waves are generated using a device called an ultrasonic disruptor or ultrasonic homogenizer. This device consists of a transducer that converts electrical energy into high-frequency mechanical vibrations.
Formation of Cavitation Bubbles: When the ultrasonic waves are transmitted to the sample, they create alternating high-pressure and low-pressure cycles. During the low-pressure cycles, small gas-filled cavities, known as cavitation bubbles, are formed within the liquid or sample. These bubbles grow rapidly during low-pressure cycles and then collapse violently during high-pressure cycles.
Cavitation Bubble Collapse: The collapse of cavitation bubbles generates intense localized energy in the form of shockwaves, microjets, and high temperatures. These physical forces result in the disruption or rupture of cells and tissues.
Mechanical and Thermal Effects: The mechanical forces generated by the collapse of cavitation bubbles cause shear stress and microstreaming within the sample. This disrupts cell membranes, breaks down cellular structures, and releases intracellular components. Additionally, the high temperatures generated during bubble collapse can also contribute to cell disruption by denaturing proteins or other heat-sensitive molecules.
Optimization of Parameters: The efficiency and effectiveness of ultrasonic disruption depend on various parameters, including the frequency and intensity of the ultrasonic waves, the duration of exposure, the sample volume, and the type of sample being disrupted. These parameters need to be optimized for each specific application to achieve the desired level of disruption without causing excessive damage or loss of biological activity.
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