USER’S GUIDE HIGH INTENSITY ULTRASONIC PROCESSOR 100 Watt Model TABLE OF CONTENTS Warranty Important Safeguards Low Surface Tension Liquids - Organic Solvents SECTION 1 – INSTALLATION Inspection Electrical Requirements Installing the Ultrasonic Processor SECTION II – OPERATION Principles of Ultrasonic Disruption Functions of Controls, Indications, and Connectors Preparations for Use Tuning Using the Ultrasonic Processor SECTION III – SERVICE INFORMATION Return of Equipment SECTION IV – OPERATING SUGGESTIONS
WARRANTY Your Ultrasonic Processor is warranted and backed by the manufacturer for a period of three years from the date of shipment against defects in material and workmanship under normal use as described in this instruction manual.
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 equipment damage. Please observe the following warnings at all times, read the operating instructions carefully before operating the equipment, and retain this instruction manual for future reference.
CAUTION LOW SURFACE TENSION LIQUIDS – ORGANIC SOLVENTS The probes (solid or with a replaceable tip) are tuned elements that resonate at a specific frequency. When working with the ½” (13mm) probe with replaceable tip, if the replaceable tip is removed or isolated from the rest of the probe, the probe will no longer resonate at the desired frequency, and the power supply will fail. Unlike aqueous (water based) solutions which rarely cause problems, solvents and low surface tension liquids are problematic.
ELECTRICAL REQUIREMENTS The Ultrasonic Processor requires a fused, single phrase 3-terminal grounding type electrical outlet capable of supplying 50/ 60 Hz at 100 volts, 115 volts, 220 volts, or 240 volts, depending on the voltage option selected. For power requirements, check the label on the back of the unit. WARNING For your personal safety, do not, under any circumstances, defeat the grounding feature of the power cord by removing the grounding prong.
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.
FUNCTIONS OF KEYS, CONTROLS, INDICATORS, AND CONNECTORS POWER SWITCH ON position – energizes the power supply. OFF position – de-energizes the power supply. Illuminates when the power supply is energized. OUTPUT CONTROL Controls the amplitude of vibrations at the probe tip. TIMER Sets the duration of ultrasonic application from 0 minute to 10 minutes. TIMER START BUTTON Illuminates when the timer or pulser is energized TIMER ON/OFF SWITCH In the ON position places the timer on standby.
PREPARATION FOR USE CAUTION Do not operate an Ultrasonic Processor that has been in a very cold or hot environment for a prolonged period of time. Wait until it has reached room temperature 1. Ensure that the AMPLITUDE is set to OFF. 2. Plug the electrical line cord into the electrical outlet. 3. If the optional footswitch is used, insert the plug into the jack located on the rear panel. Make sure that the plug is inserted forcefully all the way in. 4.
TUNING Tune the power supply in accordance with the following procedures each time a new converter or probe is used. 1. Ensure that the probe is not immersed in the liquid and that it does not come in contact with anything. 2. Set OUTPUT CONTROL TO “100”. 3. Set TIMER ON / OFF SWITCH to OFF. 4. Set PULSER to OFF. 5. Set POWER SWITCH to ON, and rotate the TUNE CONTROL clockwise or counterclockwise until minimum (not maximum) reading (usually less than 30) is obtained on the POWER MONITOR.
USING THE ULTRASONIC PROCESSOR The speed control on an automobile, can, to a certain extent, be compared to an Ultrasonic Processor. The speed control is designed to maintain the vehicles rate of travel constant. As the terrain changes, so do the power requirements. The speed control senses these requirements, and automatically adjusts the amount of power delivered by the engine in order to compensate for these ever changing conditions.
CAUTION Do not operate the power supply unless it is connected to the converter. Never allow a microtip to vibrate in air for more than 10 seconds. Do not allow the vibrating probe to come in contact with anything but the sample. 1. Ensure that the power supply is properly tuned. 2. Immerse the probe ½” (1 cm) into the liquid. 3. Set POWER SUPPLY to ON, if footswitch is used, depress footswitch.
SECTION III – 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, or that it will stop working all together. The most probable causes for malfunction are listed below and should be investigated. The unit was plugged into an electrical outlet that provides a different voltage from that required.
RETURN OF EQUIPMENT It is suggested that an Ultrasonic Processor in need of repair be sent back to the factory. In order to receive prompt service; always contact the factory before returning any instrument. Include date of purchase, model number and serial number. For instruments 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.
OPERATING SUGGESTIONS AND TECHNIQUES DISRUPTING CELLS The disruption of cells is an important stage in the isolation and preparation of intracellular products. From research levels through to production, many areas of biotechnology, particularly recombinant technology, necessitate the use of ultrasonics for cell disruption. Although some biological products are secreted from the cell or released during autolysis, many others require sonication to release intracellular material.
Microorganisms differ greatly in their sensitivity to ultrasonic disintegration. For example, the most readily disintegrated are the rod- like forms (bacilli), while the spherical organisms (cocci) are much more resistant. The group Mycobacteria, to which the tuberculosis organism belongs, is particularly difficult to disrupt. Generally, animal cells are more easily disintegrated that plant cells, and red blood cells are more readily disintegrated than muscle cells because they lack a protective cell wall.
If enzymes cannot be used, the following procedures should be considered: Freezing the sample at -70?C overnight, then thawing it in water immediately prior to ultrasonic processing. Most animal tissues can be processed fresh (unfrozen). It is important to keep fresh tissue cold and to process it quickly (within 30 minutes) after dissection. When working with fresh tissue, the cells must be sonicated immediately at the time the GITC lysis solution is added.
zymolase, glucalase and / or lyticase to produce spheroplasts that are readily lysed may also be useful. To disrupt filamentous fungi, scrape the mycelial mat into a cold mortar, add liquid nitrogen and grind to a fine powder with a pestle. The powder can then be thoroughly sonicated in lysis buffer to solubilize completely. As fungi may also be rich in polysaccharides, pretreatment with polyvinylpyrrolidone (PVP) may be beneficial.
Various methods can be used to measure the efficiency of the disruption. For example, a visual count can be made using a microscope. For greater accuracy, a protein assay could be used. This procedure is widely recognized as a good method for measuring cell disruption by taking into account the amount of protein released after disruption. The disrupted cells are then tested and checked against this number for percentage breakage. There are several types of protein assays.
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. “Sigmacote” manufactured by Sigma Chemical Co., 3050 Spruce Street, St. Louis, Missouri 63103, USA, phone (314) 771-5765, is ideally suited for that purpose. High viscosity and concentration are problematic. 2,000 cps and 15% concentration by weight are maximum limits.