USER’S GUIDE AUTOTUNE SERIES HIGH INTENSITY ULTRASONIC PROCESSOR 50 WATT MODEL • VC5020PB/VC5040PB TABLE OF CONTENTS Warranty Important Safeguards Low Surface Tension Liquids – Organic Solvents SECTION I – INSTALLATION Inspection.......................................................................................1 Electrical Requirements ................................................................1 Installing the Ultrasonic Processor ................................................
WARRANTY Your Ultrasonic Processor is warranted for a period of three years from the date of shipment against defects in material and workmanship under normal use as described in the instruction manual. During the warranty period, the manufacturer will, at its option, as the exclusive remedy, either repair or replace without charge for material and labor, the part(s) which prove to be defective, provided the unit is returned to us properly packed with all transportation charges prepaid.
IMPORTANT SAFEGUARDS READ BEFORE INSTALLING OR USING THE EQUIPMENT Your Ultrasonic Processor has been designed with safety in mind. However, no design can completely protect against improper usage, which may result in bodily injury and/or property damage. For your protection and equipment safeguard, observe the following warnings at all times, read the operating instructions carefully before operating the equipment, and retain this instruction manual for future reference.
SECTION I – INSTALLATION INSPECTION Prior to installing the Ultrasonic Processor, perform a visual inspection to detect any evidence of damage which might have occurred during shipment. Before disposing of any packaging material, check it carefully for small items. The equipment was thoroughly inspected and carefully packed before leaving our factory. The carrier, upon acceptance of the shipment, assumed responsibility for its safe delivery.
PREPARATION FOR USE 1. Mount the converter / probe assembly in a stand. Secure the clamp to the 1 ¼” (32 mm) diameter converter housing only. Do not secure the clamp to any other portion of the converter / probe assembly. 2. Connect the converter to the power supply. NOTE If the probe is not attached to the converter observe the following: a. Check for cleanliness the mating surfaces of the converter and probe, as well as the threaded stud and hole. b.
SECTION II – OPERATION PRINCIPLES OF ULTRASONIC DISRUPTION The ultrasonic power supply converts 50/60 Hz line voltage to high frequency electrical energy. This high frequency electrical energy is transmitted to the piezoelectric transducer within the converter, where it is changed to mechanical vibrations. The vibrations from the converter are intensified by the probe, creating pressure waves in the liquid.
SECTION III – USING THE ULTRASONIC PROCESSOR CAUTION 1. Do not operate the ultrasonic power supply unless it is connected to the converter 2. High voltage is present in the power supply - do not operate with the cover removed. 1. Depress the ON/OFF switch to energize the unit. The switch will illuminate. 2. Rotate the AMPLITUDE control as required. 3.
SECTION IV – SERVICE INFORMATION Your Ultrasonic Processor was designed to provide you with years of safe and dependable service. Nevertheless, because of component failure or improper usage, the possibility does exist that it might not perform, as it should, shut down due to an overload condition or that it will stop working all together. The most probable causes for malfunction are listed below and should be investigated.
RETURN OF EQUIPMENT It is suggested that if a unit is in need of repair, it should be sent back to the factory. In order to receive prompt service; always contact the factory before returning any equipment. Include date of purchase, model number and serial number. For equipment not covered by the warranty, a purchase order should be forwarded to avoid unnecessary delay. Care should be exercised to provide adequate packing to insure against possible damage in shipment.
SONICS SAFETY CERTIFICATION FORM Items being returned: Please check only one item below: ___ The equipment was never used or exposed to any radiological, biological or chemical agents and is safe to handle, use or dispose of. ___ The equipment was used but not in conjunction with or exposed to any radiological, geological or chemical agents and is safe to handle, use, or dispose of.
SECTION V – OPERATING SUGGESTIONS AND TECHNIQUES DISRUPTING CELLS The disruption of cells is an important method in the field of proteomics and in the isolation and preparation of intracellular products. Isolation of subcellular fractions and concentration of proteins allow for more efficient identification and study of proteins of interest. From research levels through to production, many areas of biotechnology, particularly recombinant technology, necessitate the use of ultrasonics for cell disruption.
Single-cell organisms (micro-organisms) consist of a semipermeable, tough, rigid outer cell wall surrounding the protoplasmic membrane and cytoplasm. The cytoplasm is made up of nucleic acid, protein, carbohydrates, lipids, enzymes, inorganic ions, vitamins, pigments, inclusion bodies, and about 80% water. In order to isolate and extract any of these substances from inside the cell, it is necessary to break the cell wall and protoplasmic membrane.
The ability to control the amplitude at the probe tip is a prerequisite for process optimization. And because each application requires its own set of processing parameters, due to variation in volume and composition, the optimum amplitude can only be determined empirically. When processing a new sample, it is recommended that the amplitude be set first at 50% (30% with a microtip) and then increased of decreased as required.
the sample remains trapped in intact cells and, therefore, is unavailable for subsequent purification. For most samples, thorough disruption can be monitored by close inspection of lysate after disruption. There should be no visible particulates, except when disrupting materials containing hard, non-cellular components, such as connective tissue or bone. When processing difficult cells, pretreatment with an enzyme to “weaken” the cell walls is beneficial.
Detergent cell lysis is sometimes used in conjunction with ultrasonic processing to facilitate disruption. Detergents break the lipid barrier surrounding cells by solubilizing proteins and disrupting lipid:lipid, protein:protein and protein:lipid interactions. Detergents, like lipids, self-associate and bind to hydrophobic surfaces.
shown to effectively release recombinant proteins located in the cytoplasm of bacteria and is recommended when lysing mammalian cells. However, this method for lysis is not recommended when working with nitrilases as it has been noticed that purified nitrilases suffer structural damages upon freezing. Most animal tissues can be processed fresh (unfrozen). However, it is important to keep them cold and to process them quickly (within 30 minutes) after dissection.
When processing a sample with ultrasonics, always immerse the probe deep enough below the surface of the sample to inhibit aerosoling or foaming, foaming substantially reduces cavitation. Processing at a lower power setting without foam is much more effective than processing at a higher power setting with foam. Decreasing the power, increasing processing time and lowering the temperature of the sample will usually prevent aerosoling and foaming. Do not use any antifoaming agents or surfactants.
Before each application, place the tip in 100% ethanol and energize the power supply for a few seconds to remove any residual substances. If still concerned about contamination from previous use, clean the probe with a disinfectant such as 20% Virkon solution and rinse with distilled water. Probes are autoclavable. To inhibit sample loss in test tube due to sticking, siliconize the test tube as follows: Wash and dry the test tube thoroughly, coat with silicone, then air dry.
Fractional protein release, Rp, is calculated using the following equation and multiplying the result by 100: Rp = Cf – Cb Ct – Cb Cf = Free protein Ct = total protein Cb = Background protein This gives the actual disruption percentage, taking into account the background levels of protein before disruption.
