USER’S GUIDE HIGH INTENSITY ULTRASONIC PROCESSOR Microprocessor Controlled 500 Watts – Model 501 600 Watt – Dual Output – Model 602 TABLE OF CONTENTS 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 INFORMA
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.
CAUTION LOW SURFACE TENSION LIQUIDS – ORGANIC SOLVENTS The probes (solid or with a replaceable tip) are tuned elements that resonate at a specific frequency. If the replaceable tip is removed or isolated from the rest of the probe, the element will no longer resonate at that 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, INDICATIONS, AND CONNECTORS FRONT PANEL TIMER Sets the duration of ultrasonic application from .1 second to 999 hours. The right hand push button selects the timing option as follows: .S=tenth of seconds, S=seconds, .M=tenth of minutes, M= minutes .H=tenth of hours, H=hours. The other three push buttons select the timing increments START BUTTON When depressed, energizes the ultrasonic. RESET BUTTON When depressed, de-energizes the ultrasonic and resets the timer.
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 POWER SWITCH 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.
No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 DESCRIPTION Converter Model CV26 Four element coupler Stepped t ip(s) 1 /8 ” (3mm) Booster Probe ½” (13mm) solid Probe ½” (13mm) with threaded end and replaceable tip Probe ¾” (19mm) solid Probe ¾” (19mm) with threaded end and replaceable tip Probe 1” (25mm) solid Probe 1” (25mm) with threaded end and replaceable tip Replaceable tip ½” (13mm) Replaceable tip ¾” (19mm) Replaceable tip 1” (25mm) Coupler Stepped tip 1/8” (3mm) Probe ½” (13mm) with threa
TUNING Tuning optimizes performance and insures maximum transfer of energy by matching the frequency of the power supply to that of converter/probe assembly. The power supply should be tuned 1) every time a new probe or accessory is used, 2) on occasions to compensate for the frequency variation caused by cavitation erosion 3) following 10 minutes of continuous operation and 4) when the sample temperature is significantly higher or lower than room temperature.
1. 2. 3. NOTE The probe is tuned to vibrate at a specific frequency – 20 kHz +/- 50 Hz. If the resonant frequency of the probe has changed, due to cavitation erosion or fracturing, minimum reading will not be obtained. If minimum reading cannot be obtained, check the instrument without the probe to determine which component might be defective. If proper tuning is obtained using the converter without the probe, the probe is defective and should be changed.
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 Never allow liquid to spill into the converter. Do not use the cup horn without a splash shield Do not allow a mic rotip or extender to vibrate in air for more than 10 seconds. When working with a microtip never allow the AMPLITUDE control to be set above the MICROTIP LIMIT “40”. Ignoring these instructions will cause the microtip to fracture. Do not allow the vibrating microtip to contact anything but the sample.
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 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.
SECTION V - 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 the n 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.