Ultrasonics Sonochemistry,DH-1200E is a multi-functional and multi-purpose instrument that uses strong ultrasound to produce cavitation effect in liquids and ultrasonically treat substances. It can be used for the fragmentation of various animal and plant cells and virus cells. Chemicals, extraction, defoaming, cleaning and accelerating chemical reactions, etc. It is widely used in biochemistry, microbiology, medicinal chemistry, surface chemistry, physics, zoology and other fields.

Ultrasonic special thermostatic reaction cup description:

The double-layer crushing cup is used in conjunction with the ultrasonic cell pulverizer. It can be connected to a low temperature thermostat to cool the system and avoid changes in the sample due to the heating of the sample during the ultrasonic process. It is more convenient and more effective than the traditional ice bath.

Main features:

●High borosilicate GG17 material, resistant to high temperature or low temperature.

●Double jacket design, the constant temperature inside the system can be achieved by entering cold water or hot water.

●There are two circulating interfaces, which are convenient for water in or out.

●Various specifications to choose from.

technical parameter:

Capacity (ml): 30 or 50, 100, 250, 500, 1000, other capacities can be customized

◆It is used for crushing animal and plant cells, bacteria, spores or tissues, extracting proteins from cells and conducting scientific culture experiments on viruses and vaccines.

◆Accelerate the reaction speed of chemistry, physics, etc. and accelerate the degassing of liquid.

◆Research and analysis of crude oil dilution, oil-water emulsification, accelerated decrystallization, glass homogenization, etc.

◆Disperse rare earths, various inorganic minerals, and prepare emulsion homogenized mixtures of nearly 1% nanometer.

◆Fast and powerful high-precision lotion for mold micro-holes and blind holes.

Purpose

1. Understand the basic principles of ultrasonic cell disruption

2. Master the basic techniques of ultrasonic cell disruption

Experimental principle

When strong sonic waves act on the solution, the generation and growth of bubbles are related to the cavitation phenomenon of fragmentation. The shock wave and scissor force caused by the cavitation phenomenon cause the cells to be lysed.

Materials and Reagents

Bacteria 10ml, beaker, shaved ice, 75% alcohol cotton ball.

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Methods of cell disruption:

1. Mechanical crushing method: refers to the use of a masher, a grinder or a homogenizer to break the cells apart.

1. High-speed tissue mashing: make the material into a thin paste liquid, place it in about 1/3 of the volume of the cylinder, close the cylinder lid tightly, turn the governor to the slowest position first, turn on the switch, and gradually accelerate to the desired position. speed required. This method is suitable for animal visceral tissue, plant succulent seeds, etc.

2. Homogenization by glass homogenizer: first place the chopped tissue in a tube, then insert the grinding rod into the tube to grind it back and forth, and move it up and down to grind the cells. High, suitable for small amount and animal organ tissue.

2. Physical fragmentation method: refers to the use of temperature difference, pressure difference or ultrasonic wave to break cells apart.

1: Treat the cell suspension with a certain power of ultrasonic wave, so that the cells are violently shaken and broken (the cell wall and organelles are broken by the vibration force of the ultrasonic wave).

Mechanism: It may be related to the cavitation phenomenon of bubble generation, growth and fragmentation when strong sonic waves act on the solution, and the shock wave and scissor force caused by the cavitation phenomenon cause cell lysis.

The efficiency of sonication depends on the sound frequency, sound energy, treatment time, cell concentration and cell type, etc. (Pay attention to cooling down when using to prevent overheating).

2. High-pressure fragmentation: The cell suspension is ejected from the annular gap of the high-pressure chamber onto the stationary impact ring, and is forced to change direction and flow out through the outlet tube. During this process, the cells undergo high-speed shear collisions and changes from high pressure to atmospheric pressure, thereby breaking up and releasing the contents.

This is an ideal method for gentle, thorough cell disruption.

3. Repeated freeze-thaw method: freeze the cells below -20 degrees, thaw at room temperature, and repeat several times. Due to the formation of ice particles in the cells and the increase of the salt concentration of the remaining cell fluid, the swelling will be caused, and the cell structure will be broken.

3. Chemical fragmentation method: refers to the use of formaldehyde, acetone and other organic solvents or surfactants to act on the cell membrane, so that the structure of the cell membrane is damaged or the permeability is changed.

Some animal cells, such as tumor cells, can be destroyed by the use of sodium dodecyl sulfonate (SDS), sodium deoxycholate and other cell membranes. The concentration is generally 1 mg/ml.

4. Enzymatic fragmentation method: Use appropriate enzymes to destroy the cell wall, and then fragment the protoplasts in a hypotonic solution.

Bacterial cell walls are thicker and can be treated with lysozyme for better results.

Standard formulation of lysate: : 50mM Tris-HCl (pH8.5~9.0), 2mM EDTA, 100mM NaCl, 0.5% Triton X-100, 1mg/ml lysozyme. (Lysozyme works better in this pH range).

Comprehensive description: No matter which method is used to disrupt tissue cells, intracellular proteins or nucleic acid hydrolase will be released into the solution, which will biodegrade macromolecules, resulting in a reduction in the amount of natural substances. Adding diisopropyl fluorophosphoric acid (DFP) Can inhibit or slow down autolysis; the addition of iodoacetic acid can inhibit the activity of those proteolytic enzymes that require a sulfhydryl group in the active center, and the addition of phenylmethanesulfonyl fluoride (PMSF) can also eliminate the activity of proteolytic enzymes, but not all, And it should be added several times while crushing; in addition, pH, temperature or ionic strength can also be selected to make these conditions suitable for the extraction of the target substance.

Precautions before use:

1. Select an appropriate probe according to the volume of the sample to be processed. Before and after use, the probe must be wiped with an alcohol cotton ball, and the probe must be replaced with a special wrench;

2. When the AMPLITUDE knob is turned on, the probe must not be exposed to the air, and in special cases (test probe) should not exceed 10 seconds;

3. Prepare an ice box before use, and the sample must be placed in an ice bath during processing, and the sample concentration should not be too large.

Ultrasonics Sonochemistry DH-1200E is a multi-functional and multi-purpose instrument that uses strong ultrasound to produce cavitation effect in liquids and ultrasonically treat substances. It can be used for the fragmentation of various animal and plant cells and virus cells. Chemicals, extraction, defoaming, cleaning and accelerating chemical reactions, etc. It is widely used in biochemistry, microbiology, medicinal chemistry, surface chemistry, physics, zoology and other fields.