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250 - 450 Sonifier Analog Cell Disruptor User`s Manual

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1. 2 4 tab 6 1 Temperature Rise Variations 6 5 tab 6 2 Horn Amplitudes 6 8 tab 7 1 Symptom Probable Cause 7 7 tab B 1 Replacement Parts B 1 tab D 1 Accessories Parts List D 1 100 413 016 Rev C vii BRANSON viii 100 413 016 Rev C 250 450 Sonifier Chapter 1 Safety and Support User s Manual Chapter 1 Safety and Support 1 1 Safety Considerations 1 2 1 2 About this Manual 1 3 1 3 Intended Use of the System 1 4 1 4 Regulatory Compliance 1 4 1 5 Warranty 1 5 1 6 How to Contact Branson 1 6 1 6 1 Before Calling Branson for Assistance 1 6 1 7 Returning Eguipment for Repair 1 7 1 8 Obtaining Replacement Parts 1 9 100 413 016 Rev C 1 1 Safety Considerations BRANSON 1 1 Safety Considerations Observe the following safety considerations when operating the Analog Sonifier 1 Make sure that the equipment is properly grounded DO NOT oper ate if it is not 2 Make sure the unit is equipped with a three conductor cord and three prong grounding type plug and must be plugged into a three prong ground type wall receptacle DO NOT under
2. D 2 5 1 2 Mechanical Specifications 5 3 5 1 3 Dimensions 5 4 100 413 016 Rev C 5 1 Chapter 5 Technical Specifications BRANSON Specifications 5 1 Specifications 5 1 1 Electrical Specifications Table 5 1 Electrical specifications Controls amp Timer 0 15 Minutes and Hold Displays Output control Duty Cycle Continuous ON OFF Switch Pilot Light Output control Adjusts amplitude of power supply output voltage Range 10 100 of nominal converter amplitude Duty Cycle Intermittent pulse duration adjustable 10 90 1 pulse per second Continuous Processing Time Connector RF connects 20kHz high voltage to converter con nections Model 102C Horn 19 850 20 050 KHz Frequency Line Voltage Model 250 100 120V 50 60 Hz 2 0 A 200 245V 50 60 Hz 1 2 A Model 450 100 120V 50 60 Hz 4 0 A 200 245V 50 60 Hz 2 3 A Fuse 5A 250V 5x20mm SB or TYPET Leakage 0 5mA Maximum 115V line to ground Power Model 250 150 Watts Minimum Model 450 350 Watts acceptable output power 100 413 016 Rev C 250 450 Sonifier Chapter 5 Technical Specifications User s Manual Specifications Table 5 1 Electrical specifications Minimum Model 250 120 Watts Starting Power Model 450 300 Watts Temperature Operating 41 F 5 C 104 F 40 C Ambient Storage and Shipping 20 F 29 C to 158 F 70
3. 7 4 7 4 2 Stud Replacement 7 6 7 4 3 Horn Tip Cleaning 7 7 7 4 4 System Trouble Analysis Chart 7 7 ii 100 413 016 Rev C 250 450 Sonifier User s Manual Appendix A Applications Appendix B Parts List Appendix C Accessories C 1 Micro Tip C 1 C 2 Cup Horn ee ee C 2 C 3 Flow thru Horn C 3 C 4 Rosette Cell C 3 C 5 Continuous Flow Attachment C 4 C 6 Soundproof Box C 4 Appendix D Accessories Parts List D 1 Accessories Parts List D 1 100 413 016 Rev C iii BRANSON 100 413 016 Rev C 250 450 Sonifier User s Manual fig fig fig fig fig fig fig fig fig 100 413 016 Rev C 2 1 2 2 2 3 4 1 4 2 4 3 5 1 5 2 C 1 List of Figures Model 250 450 Sonifier Main Components2 2 Power Supply Front Panel2 4 Power Supply Rear Panel2 6 Converter Horn Assembly in sound proof enclosure 4 3 Methods for Connecting Removing Horn from Converter4 5 Connecting or Removing Tip from Horn4 6 250 450 Sonifier Side View Dimensions5 4 250 450 Sonifier Front View Dimensions5 4 Soundproof BoxC 4 vi BRANSON 100 413 016 Rev C 250 450 Sonifier User s Manual List of Tables tab 2 1 Front Panel Device and Function
4. 1 2 1 2 About this Manual 1 3 1 3 Intended Use of the System 1 4 1 4 Regulatory Compliance 1 4 1 5 Warranty 1 5 1 6 How to Contact Branson 1 6 1 6 1 Before Calling Branson for Assistance 1 6 1 7 Returning Eguipment for Repair 1 7 1 8 Obtaining Replacement Parts 1 9 Chapter 2 Introduction 2 1 Main Components 2 2 2 2 Principle of Operation 2 2 2 3 250 450 Sonifier Power Supply Front Panel 2 4 2 4 Model 250 450 Sonifier Power Supply Rear Panel 2 6 Chapter 3 Unpacking and Handling 3 1 Unpacking and Handling 3 1 Chapter 4 Installation and Setup 4 1 Placement of Equipment 4 2 4 2 Power Requirements 4 2 4 3 Setup Procedure 4 3 4 4 Testing the Equipment 4 4 4 5 Connecting and Removing Tips Horns and Converters 4 5 4 5 1 Connecting Horn to Converter 4 5 4 5 2 Connecting Tip to Horn 4 5 4 6 Guards and Safety Equipment 4 7 100 413 016 Rev C i BRANSON Chapter 5 Technical Specificati
5. 1 10 100 413 016 Rev C 250 450 Sonifier Chapter 2 Introduction User s Manual Chapter 2 Introduction 2 1 Main Components 2 2 2 2 Principle of Operation 2 2 2 3 250 450 Sonifier Power Supply Front Panel 2 4 2 4 Model 250 450 Sonifier Power Supply Rear Panel 2 6 100 413 016 Rev C 2 1 Main Components BRANSON 2 1 Main Components The main components of the 250 450 sonifiers are the power supply the converter and the horn shown in Figure 2 1 below Figure 2 1 Model 250 450 Sonifier Main Components 2 2 Principle of Operation The power supply converts AC line voltage to 20kHz electrical energy This high frequency electrical energy is fed to a converter where it is converted to mechanical vibrations The heart ofthe converter is a lead zirconate titanate electrostrictive element which when subjected to an alternating voltage expands and contracts The converter vibrates in a longitudinal direction and transmits the motion to the horn tip immersed in the solution which causes cavitation The implosion of microscopic bubbles or cavities in the solution results causing the molecules in the medium to become intensely agitated The Sonifier functions in two modes Pulsed and Continuous In Pulsed Mode ultrasonic vibrations are transmitted to a solution at a rate of one pulse per second This pulse duration can be adjusted from 0 1 to 0
6. 6 6 6 2 5 Disrupting Tissues and Solids 6 6 6 2 6 Using Glass Powders with Solution 6 7 6 3 Horn and Micro Tip Amplitudes 6 8 100 413 016 Rev C 6 1 General Operation BRANSON 6 1 General Operation In general starting conditions are as follows 6 1 1 Timed Operation Turn Timer Control clockwise to the desired processing time Set Duty Cycle Control to Constant and Output Control to 1 With the tip of the horn immersed about 1 4 to 1 2 inch in the solution to be pro cessed turn power switch to ON To increase ultrasonic intensity turn Output Control clockwise to the next highest number pausing at each setting in order to evaluate results until the optimum setting is reached 6 1 2 Continuous Operation Turn Timer Control fully counterclockwise to the Hold position Set Duty Cycle Control to Constant and Power Switch to On Adjust Output Control to the desired required intensity 6 1 3 Pulsed Operation Use of the Duty Cycle Control enables the ultrasonic energy to be pulsed one pulse per second The control settings from 10 to 90 changes the duration of the pulse from 10 to 90 of each second respectively Pulsing the ultrasonic energy retards the rate of temperature increase in the solution that results from the ultra sonic activity To operate in the pulsed mode set the Timer Control to Hold ifthe process is not to be timed or a processing t
7. 5 Notify Branson prior to shipping the equipment To return equipment to Branson you must first obtain an RGA number from Branson or the shipment may be delayed or refused 6 For equipment not covered by warranty include a purchase order for the repair costs to avoid delay 7 Pack carefully in original packing material to avoid damage in ship ment 8 Return general repairs by any desired method Send priority repairs by air freight 9 Prepay the transportation charges FOB Laredo Texas U S A Notes 1 8 100 413 016 Rev C 250 450 Sonifier Chapter 1 Safety and Support User s Manual Obtaining Replacement Parts 1 8 Obtaining Replacement Parts You can reach Branson Parts Store at the following telephone numbers Branson Part Store direct telephone number 877 330 0406 fax number 877 203 0404 Many parts can be shipped the same day if ordered before 2 30 p m Eastern time A parts list is in Appendixes B and D ofthis manual listing descriptions and EDP part numbers If you need replacement parts coordinate the following with your purchasing agent Purchase order number Ship to information Bill to information e Shipping instructions air freight truck etc Any special instructions for example Hold at the airport and call Be sure to give aname and phone number Contact name information 100 413 016 Rev C 1 9 Obtaining Replacement Parts BRANSON
8. into electrical outlet 3 Line filter has failed Fan does not operate Indicator light 1 Unplugged power supply fails to illuminate when power sup 2 Line cord has failed ply is energized 3 Power Switch has failed 4 Fuse F1 has failed 5 Line filter has failed Fan motor has failed Line cord failed Power Switch has failed Fuse F1 has failed Line filter has failed Fan does not operate when power supply is energized Indicator light illuminates ge dt 100 413 016 Rev C 7 7 Chapter 7 Maintenance Reconditioning the Stack Interface BRANSON Table 7 1 Symptom Probable Cause Symptom Probable Cause Fuse F1 fails when ON OFF switch 1 Underrated fuse Fl is energized 2 Incorrect voltage 3 Fan motor has failed 4 Power Supply module has failed Ultrasonic power is delivered to the 1 Meter has failed horn No indication on meter No or inconsistent ultrasonic power 1 Foreign material between horn sur Some indication on meter BE A A face and replaceable tip Loose or worn out tip Horn is loose or failed Horn stud is loose or failed Converter cable has failed Converter has failed If used with a treatment chamber horn tip may not be immersed Little or no meter indication when ON OFF switch is energized Fan operates a gt U Timer has not been set has com pleted cycle or has failed Thermal switch on power supply modul
9. If you have any questions please contact your Branson representative The product war ranty information is summarized below Warranty When used in accordance with written instructions and under normal operating conditions Branson Ultrasonics Corp equipment is guaranteed to be free of defects in MATERIAL and WORKMANSHIP for two 2 years from the date of original delivery by BRANSON or by an authorized representative Any unit which proves defective during the stated period will be repaired free of charge or replaced at the sole discretion of Branson Ultrasonics Corp F O B Laredo Texas or an authorized repair station as advised by BRANSON provided the defective unit is returned properly packed with all transportation charges prepaid A limited warranty as specified may apply to certain components of the equipment Warranty Exceptions This warranty shall not apply to equipment subjected to misuse improper instal lation alteration neglect accident or improper repair This warranty is limited to the original purchaser and is not transferable Horns and tips fabricated by Branson for use in equipment described in this man ual are manufactured to exacting parameters Using altered or modified horns and tips or horn and tips otherwise unqualified by Branson can produce undue stresses that may damage the equipment Equipment failures resulting from using unqual ified horns and tips are not covered by the Branson warranty Microt
10. packed column Clostridium quickly disrupts all types Coagulase globulin ultrasonic treatment before precipitation yields signifi cantly more enzyme 100 413 016 Rev C A 3 Appendix A Applications BRANSON Collagen an excellent fragmentation Colletotrichum capsici spores 5 ml with 6 million spores ml completely disrupts in 4 minutes Cortiscosteroid particle size can be reduced to approximately 5 microns while large volumes can be treated at the rate of approximately 30 ml minute on a con tinuous flow basis Corynebacterium completely disrupts in 5 minutes with 50 protein release and excellent enzyme activity Crypstostroma corticale maple bark spores a concentrated solution completely disrupts inl4 minutes Cryptococcus laurentii completely disrupts in 7 minutes with good protein release and enzyme activity Crystal reduction large crystals of an organic compound suspended in iso propanol can be reduced in diameter 10 40 Cyanidium caldarium a concentrated solution completely disrupts in 6 min utes Dental plaques 5 ml solution concentration 1 to 10 000 low power setting 53 500 000 organisms per ml were obtained in 45 seconds Desullovibrio vulgaris less than 30 seconds of ultrasonic treatment resulted in release of TCA enzymes Diplococcus completely disrupts in 5 minutes DNA breaks chains on low power instantly Controlled degradation may be obtained Dyes excellent rapid dispers
11. Cell The Rosette Cell made of borosilicate glass has a conical shape with three side arms through which the solution 1s driven by the ultrasonic horn thereby expos ing the substance to ultrasonic energy repeatedly during circulation When the Rosette Cell is immersed in a cooling bath the enlarged glass surface area plus circulation through the side arms provide an efficient means of heat exchange The Cell is available in three sizes 35 300 ml 25 180 ml and 8 25 ml 100 413 016 Rev C C 3 Appendix C Accessories BRANSON C 5 Continuous Flow Attachment This attachment which is screwed on to the disruptor horn permits continuous processing of low viscosity materials at rates up to 38 liters per hour Designed primarily for emulsifying dispersing and homogenizing the attachment will dis rupt most cells with the exception of the more difficult species The materials being treated may be passed through the attachment more than once to obtain desired results For cooling a water jacket with input output and overlow con nections is provided C 6 Soundproof Box Even though ultrasound is above the audible range ofthe human ear mechanical noise occurs when liquids are treated ultrasonically The soundproof box reduces this noise by 25 to 28 db It is especially useful when the Sonifier is used for extended periods of time Figure C 1 Soundproof Box Sny Va C 4 100 413 016 Rev C 250 450 Sonifier Instru
12. any circum stances remove the power cord ground prong 3 DO NOT operate equipment with the cover removed High voltage is present within the equipment when connected 4 DONOT allow the horn or micro tip to contact lab stands beakers etc or horn micro tip failure may result 5 DO NOT turn on the ultrasonics without the converter and tool attached 6 DO NOT touch the horn or tip when ultrasonics are active When handling removing or attaching a tool or tip be sure that the ON OFF switch is set to OFF Touching the tool while the unit is on can result in serious personal injury If the Sonifier is used in a manner not specified by Branson the protec tion provided by the equipment may be impaired 1 Sonifier is a registered trademark of Branson Ultrasonic Corporation 1 2 100 413 016 Rev C 250 450 Sonifier Chapter 1 Safety and Support User s Manual About this Manual 1 2 About this Manual This manual contains operation and service instruction for the Sonifier Model 250 450 It contains background information essential to the proper use and care of this equipment The following meaning applies to signal words found throughout this manual WARNING indicates a hazardous situation which if not avoided could result in death or serious injury CAUTION indicates a hazardous situation which if not avoided could result in minor o moderate injury Notice is used to address recommendeg practices not related t
13. can be evenly dispersed giving a quick representative sample of calls for cytologic examination Cells are liberated from the mucous of sputum that had been immersed in 50 alcohol or a fixative Ragweed pollen 15 ml dispersion completely disintegrates in 11 minutes Rat bone 1 2 gm disintegrates in 4 minutes Rat liver complete disruption in 3 minutes Rat liver mitochondria ultrasonic treatment has been used extensively for the varied research performed on this material Disruption occurs in seconds Rat skin 1 gm completely disintegrates in 4 minutes Red and white blood cells ultrasonic treatment breaks particle size to 100 Angstroms Complete disruption in 1 minute 25 gms 100 ml saline or plasma sample treated 15 seconds 35 disruption Adenosine triphosphate was shown to be membrane bound by this method Reovirus dissociates cell bound and aggregated virus Maximum titer with 4 ml of virus was achieved in 2 minutes Retinal outer segments ultrasonic treatment breaks particles down to almost molecular size Rhodopseudomonis palustris complete disruption in 4 minutes Rhodospirillum rubrum excellent disruption in a few seconds Rimosus Monocellular elements from surface grown colonies obtained in 1 minute Complete disruption in 5 minutes 50 disruption in 2 minutes RNA rapid and thorough re suspension of 9 TCA pellets during extractions Rocks excellent for disaggregation of sedimentary rock and for cleaning
14. disaggregation of all fine grained sedimentary rocks Shellfish by drilling a clean hole with the micro tip various fluids or samples may be withdrawn or injected from living shellfish without destroying the ani mals Shigella quick disruption Skin 1 gm disintegrates in about 4 minutes Epidermal homogenates that respire and utilize substrate can be extracted Soil separates solid particles without use of oxidants acids or peptizing agents and yields stable suspensions Sperm human tails are broken instantly Heads are broken in 20 minutes Sputum cancer cells are more easily detected after ultrasonic treatment due to even dispersion of cells and sputum and complete liberation of the cells from spu tum Staphylococcus a concentrated solution disrupts 98 in 10 minutes With 1 gm cells wet wt to 2 gm water 54 5 mg ml of protein was released Starch obtained by extracting from green plant leaf homogenate Streptococcus Group A a 20 suspension in a 15 ml solution completely disrupts in 15 minutes A 10 100 413 016 Rev C 250 450 Sonifier Appendix A Applications Instruction Manual Streptomyces monocellular elements from surface grown colonies obtained in 1 minute Complete disruption in 5 minutes 50 disruption in 2 minutes Subcellular particles may be separated or broken depending on power selec tion and length of time Sulfanilamide excellent dispersion in less than 1 minute Continued ultrasoni
15. minute Complete disruption in 5 minutes 50 disruption in 2 minutes Antigen the Sonifier Cell Disruptor is used extensively to produce antigens and vaccines It can increase yield or expose otherwise unobtainable sites Aorta 1 gram disintegrates in 2 minutes Aphanomyces after blending complete disruption in 3 minutes Arthobacter tumescens 10 gm in 40 ml in 5 minutes for O coumaric reduc tose Ascaris eggs 8 ml concentrated solution completely disrupts in 4 minutes Asperigillus complete disruption in 4 minutes Aurefaciens monocellular elements from surface grown colonies obtained in 1 minute Complete disruption in 5 minutes 50 disruption in 2 minutes 100 413 016 Rev C A 1 Appendix A Applications BRANSON Azotobacter vinelandii 15 ml buffered solution 200 mg wet wt per ml com pletely disintegrates in 2 minutes B anthracis 80 disruption of anthracis in 4 minutes Complete disruption of 10 ml of eryisipelothrix rhusipathiae in 10 minutes B cereus veg cells disruption in a few seconds B cereus spores disruption of 10 mg 6 ml in 13 minutes B megaterium spores complete breakage ofa concentrated 6 ml solution in 15 minutes B sphaericus major disruption in 1 3 minutes B stereothermophilis spores complete disruption in 2 minutes B subtilis disruption of 5 gm wet wt 15 ml buffer in 5 minutes B subtilis veg cells heavy suspension clears in 1 minute Bacillus stereothermophulus th
16. of Equipment 4 2 4 2 Power Requirements 4 2 4 3 Setup Procedure 4 3 4 4 Testing the Equipment 4 4 4 5 Connecting and Removing Tips Horns and Converters 4 5 4 5 1 Connecting Horn to Converter 4 5 4 5 2 Connecting Tip to Horn 4 5 4 6 Guards and Safety Equipment 4 7 100 413 016 Rev C 4 1 Placement of Equipment BRANSON 4 1 Placement of Equipment Install the unit in an area away from radiators or heating vents A fan maintains safe operating temperatures in the power supply by circulating air over the components Therefore place the power supply so that the air intake located at the rear of the power supply is not blocked If the temperature within the unit becomes excessive a thermal cutout switch will disconnect the power and keep it disconnected until the ambient temperature cools to a safe operating level Although the unit will automatically be re energized when a safe operating temperature has been reached it should be checked to determine the reason for the initial cutout Periodically unplug the unit and clean the air intake at the rear of the power supply to ensure that dust or dirt is not restricting the flow of air 4 2 Power Requirements The Sonifier unit requires a single phase three wire 117V or 200 245V
17. once or twice in the same direction Rotate the part 120 degrees placing your thumb over the span ner wrench hole and lap the part the same number of times as described above 10 Rotate the part another 120 degrees to the next spanner wrench hole and repeat the lapping procedure O ONO Re examine the mating surface If necessary repeat steps 5 through 10 until you remove most of the contaminant 100 413 016 Rev C 7 5 er Interface BRANSON 7 4 2 Stud Replacement The Stud is intended to be a use only once part as it has knurls on its end which bite into the relatively softer horn material The studs are also specially designed to withstand ultrasonic stresses If you must replace the stud it is very important to clean the threads and the horn of the previous shavings 1 Using a clean cloth or towel clean the threaded hole 2 Examine the threaded hole for stripped threads Do not use a damaged ultrasonic horn or converter Threaded studs cannot be reused in titanium horns Clean the threaded hole before inserting the new stud Apply one drop of Loctite threadlocker to the stud and insert it into the horn 5 Tighten the stud Use the following torque specifications Stud Size Torque Specification Stud EDP number 3 8 24 x 11 4 in 290 in Ibs 33 Nm 100 098 121 3 8 24 x 11 2 in 290 in Ibs 33 Nm 100 098 120 1 2 20 x 11 4 in 450 in Ibs 51 Nm 100 098 370 1 2 20 x 11 2 in 450 in Ibs 51 Nm 1
18. 00 098 123 1 2 20 to 3 8 24 x lin 250 in Ibs 28 Nm 100 098 249 Adaptor stud After the new stud has been inserted you can reassemble the Horn to the Con verter Follow the same procedure as found in Section 4 9 Connecting and Removing Tips Horns and Converters beginning on page 4 5 1 Loctite is a registered trademark of Henkel Corporation U S A 7 6 100 413 016 Rev C 250 450 Sonifier Chapter 7 Maintenance User s Manual Reconditioning the Stack Interface 7 4 3 Horn Tip Cleaning Take the following steps to clean the horn s tip threads 1 If the horn has a replaceable tip remove it and clean its threads in alcohol 2 Swab out the end of the horn threads with a cotton swab and alco hol 3 Make sure both the horn and the tip are clean and completely dry before you reassemble them For information on tightening the tips see Section 4 5 Connecting and Removing Tips Horns and Converters beginning on page 4 5 4 Use a spanner wrench on the horn and an open end wrench on the tip to install the tip using the following torque specifications 1 4 20 tighten at 90 inch Ibs 10 16 Newton meters 3 8 24 tighten at 180 inch Ibs 20 33 Newton meters 7 4 4 System Trouble Analysis Chart Table 7 1 Symptom Probable Cause Symptom Probable Cause Main power fuse fails or circuit Line cord is shorted reader trips when system is plugged 2 Power Switch has failed
19. 2 4 Model 250 450 Sonifier Power Supply Rear Panel Figure 2 3 Power Supply Rear Panel BRANSON Table 2 2 Rear Panel Device and Function Item Function 1 Power Cord Connects the power supply to an electrical outlet 2 Fuse 4 AMP Overload Protection 3 RF Cable Connects the power supply to the converter 100 413 016 Rev C 250 450 Sonifier Chapter 3 Unpacking and Handling User s Manual Unpacking and Handling Chapter 3 Unpacking and Handling 3 1 Unpacking and Handling 3 1 3 1 Unpacking and Handling Unpack the model 250 450 Sonifier unit as soon as 1t arrives Normal precautions in unpacking and reasonable care in handling should be exercised to avoid possi ble damage to the unit Visually inspect all external controls indicators and sur faces to detect any damage which might have occurred during shipment The shipping company is responsible for damage to equipment during shipment If damage has occurred notify the shipping company immediately to establish proper basis for claim In case of damage do not discard packing material until shipping com pany has inspected for cause of damage 100 413 016 Rev C 3 1 Unpacking and Handling BRANSON 3 2 100 413 016 Rev C 250 450 Sonifier Chapter 4 Installation and Setup User s Manual Chapter 4 Installation and Setup 4 1 Placement
20. 250 450 Sonifier Analog Cell Disruptor User s Manual EDP 100 413 016 Rev C BRANSON Ultrasonics Corporation 41 Eagle Road Danbury Connecticut 06813 1961 U S A 203 796 0400 http www sonifier com BRANSON 100 413 016 Rev C 250 450 Sonifier User s Manual Manual Change Information At Branson we strive to maintain our position as the leader in ultrasonic plastics Joining cleaning and related technologies by continually improving circuits and components in our equipment These improvements are incorporated as soon as they are developed and thoroughly tested Information concerning any improvements will be added to the appropriate man ual section s at the next printing Therefore when requesting service assistance for specific units refer to the printing date which appears in the lower right corner of this page Copyright Copyright 1998 by Branson Ultrasonics Corporation Danbury Con necticut U S A All rights reserved Sonifier is a registered trademark of Branson Ultrasonics Corporation Danbury Connecticut U S A Loctite is a registered trademark of Henkel Corporation U S A Other trademarks and service marks respectfully mentioned herein are held by their respective owners BRANSON Ultrasonics Corporation January 2011 BRANSON 100 413 016 Rev C 250 450 Sonifier User s Manual Table of Contents Chapter 1 Safety and Support 1 1 Safety Considerations
21. 50 60 Hz source The 117V line cord is equipped with a NEMA 5 15P plug and requires a NEMA 5 15R receptacle The 200 245V line cord for European units is equipped with a CEE 7 7 plug To prevent the possibility of electrical shock always plug the Sonifier unit into a grounded power source Setup Procedure 4 2 100 413 016 Rev C 250 450 Sonifier Chapter 4 Installation and Setup User s Manual Setup Procedure 4 3 Setup Procedure Figure 4 1 Converter Horn Assembly in sound proof enclosure a Set ON OFF switch to OFF b Plug the line cord intone appropri ate electrical power outlet ensuring that the power supply is grounded to prevent possibility of an electrical shock c Ensure that the horn is screwed into the converter and tightened ade quately Remember to use washer in between Horn and Converter The recommended torque is 220 inch pounds 24 85Nm d Mount converter horn assembly in a laboratory stand or Branson sound enclosure as shown in Figure 4 1 Secure clamp on converter e Ensure that the tip is screwed into the horn and tightened adequately Recommended torque is 90 in Ibs 10 16Nm for a flat tip with 1 4 20 threads To remove a horn use the spanner wrenches supplied See Section 4 5 Connecting and Removing Tips Horns and Converters 100 413 016 Rev C 4 3 Testing the Eguipment 7 BRANSON 4 4 Testing the Eguipment To establish a base line for future testing to verify 1f the e
22. 6 Rev C D 3 Appendix D Accessories Parts List Table D 1 BRANSON Accessories Parts List ITEM BRANSON DESCRIPTION EDP No Double Step Micro Tip Assembly Stud Rosette Cool ing Cell A two piece horn consisting of a coupling sec tion and a lower tip The standard disruptor horn must be removed prior to using this tip recommended for use on extremely small vol umes 0 5 20ml applications include red and white blood cells tissue culture cells Hela cells Overall length is 9 1 8 with 1 8 dia in the lower 2 1 8 The double step microtip is to be used only with coupler 101 147 050 Kit Coupler amp Micro Tip 101 063 212 Coupler Section Only 101 147 050 Micro Tip Section Only 101 148 063 Replacement adaptor stud 100 098 249 Horn to Converter Borosilicate glass cell has conical shape with three arms to allow circulation of substance being processed When the cell is immersed in a cooling bath the enlarges glass surface areas plus circulation through the arms pro vide and effective means of heat exchange Model 25 8 25 ml 201 123 001 Model 50 25 180ml 201 123 002 Model 250 35 300ml 201 123 003 100 413 016 Rev C 250 450 Sonifier Instruction Manual Table D 1 Appendix D Accessories Parts List Accessories Parts List ITEM DESCRIPTION BRANSON EDP No Tissue Disrup tor Soundproof Enclosure Designed for disint
23. 9 per second enabling 2 2 100 413 016 Rev C 250 450 Sonifier Chapter 2 Introduction User s Manual Principle of Operation a solution to be processed at full ultrasonic intensity while limiting temperature build up especially valuable when processing heat volatile solutions and temperature sensitive material In Continuous Mode the Sonifier can be set up for continuous duty The 200 and 400 Watts output power specified for the Models 250 and 450 respectively is their maximum power capability The actual power developed for any application will depend upon the load on the horn face The Sonifier is a constant amplitude device i e as the load or pressure on the horn face increases the power supply will develop more power to maintain the amplitude for any given output control setting When the horn is operated in air the horn is subjected to minimum pressure and minimum power The load will increase when the horn is immersed in a liquid the more viscous the liquid the higher the load and the more power required If a flow through cell which can be pressurized is used thereby applying pressure on the horn even more power will be required For any given application more power will result when a horn of higher amplitude or with larger radiating surface is used or when any horn is driven at higher amplitude by increasing the output control setting System protection constantly monitors various operating parameters of the pow
24. C Tuning Factory set no tuning necessary Output control at maximum setting 5 1 2 Mechanical Specifications Table 5 2 Mechanical specifications Converter Weight 4 Ibs 1 8 kg with horn Dimensions 7 L x 2 1 2 dia 180 x 60 mm Horn Tip Diameter 1 8 1 3 2 25mm dia depending on process needs Power Weight 15 Ibs 6 8 kg Supply 100 413 016 Rev C 5 3 Chapter 5 Technical Specifications Specifications BRANSON 5 1 3 Dimensions Figure 5 1 250 450 Sonifier Side View Dimensions 1 75 B 11 19 gt lt 16 83 18 96 gt Figure 5 2 250 450 Sonifier Front View Dimensions le 7 38 gt SONIFIER 9 625 put Comm 5 4 100 413 016 Rev C 250 450 Sonifier Chapter 6 Operation User s Manual Chapter 6 Operation 6 1 General Operation 6 2 6 1 1 Timed Operation 6 2 6 1 2 Continuous Operation 6 2 6 1 3 Pulsed Operation 6 2 6 2 Operating Considerations 6 3 6 2 1 Limiting Temperature Rise 6 3 6 2 2 Vessel Capacity and Speed of Temperature Rise 6 5 6 2 3 Minimizing Undesirable Factors 6 5 6 2 4 Sterilizing and Preventing Cross Contamination
25. Cause on page 7 7 to locate the symptom that most clearly describes your problem 7 2 1 Tip Erosion Horn tips can erode Tip erosion is a side effect of the cavitation process that occurs when liquids are exposed to ultrasonic energy The rate of erosion depends on the intensity of power applied the corrosiveness of the liquid being treated and the amount of use Periodically inspecting the tip will help you recognize erosion early As erosion progresses the color of the tip changes from its original polished appearance first to light grey and then to dark grey Concentric rings begin to appear and finally the tip becomes rough and pitted resulting in loss of power output As it erodes the tip can also introduce metal particles into the solution causing it to darken or discolor Eventually erosion may become significant at which point the tip will require replacement 7 2 2 Power Output Loss If you observe a loss in power several factors can be the cause Ensure that the tip is not corroded and is polished as noted in the preceding paragraph If the horn has a replaceable tip remove the tip and clean its threads in alcohol 7 2 100 413 016 Rev C 250 450 Sonifier Chapter 7 Maintenance User s Manual Preventive Maintenance Place the horn in a container of alcohol and turn the power on for a few seconds to clean the interior threads Remove the horn from the alcohol Operate the horn in air for a few seconds to dry t
26. Flow Attachment C 4 C 6 Soundproof Box C 4 Special disruptor horns various horn tips and a variety of vessels for batch or con tinuous processing are available for use with Branson Sonifiers Accessories include the following C 1 Micro Tip Particularly useful for processing small volumes micro tips are available in two designs tapered and double stepped to meet your application reguirements The tapered micro tip attaches directly to a standard 1 2 tapped disruptor horn The amplitude at the end of a tapered tip 1s three and a half times greater than that of the standard horn The tapered tip is recommended for difficult applications such as spores fungi yeast muscle and connective tissue Excellent results can be achieved on volumes ranging from 3 to 20 ml in a comparatively short period of time The diameter of the tapered micro tips are 1 8 3 2 mm 3 16 4 8mm and 1 4 6 4mm 100 413 016 Rev C C 1 Appendix C Accessories BRAN S ON The double stepped micro tip is a two piece horn consisting of a coupling section and a lower tip Because the coupling section 1s attached directly to the converter the standard disruptor horn must be removed prior to using the double stepped micro tip Recommended for use on extremely small volumes the double stepped tip can be used to treat volumes ranging from 0 5 to 20 ml Applications for this tip include red and white blood cells tiss
27. apter 6 Operation User s Manual Operating Considerations 6 2 2 Vessel Capacity and Speed of Temperature Rise The smaller the volume the more difficult the cooling procedure becomes For example using any given power input to treat 5 ml for a long period would require a cooling bath of approximately 35 C to maintain the sample at or below 5 C In comparison the processing of 200 ml would require a cold bath of only 0 C Table 6 1 shows typical temperature rises for sample sizes of 25 ml and 100 ml using a 250 Sonifier A polyethylene container was used with a 1 2 12 7 mm diameter probe with a probe depth of 1 2 inch 12 7 mm and a starting tempera ture of 25 Average Diff was the average difference among duplicate runs Table 6 1 Temperature Rise Variations Sample Size 25 ml 100m l Power Level 3 7 10 3 7 10 Seconds Temperature after sonic time 30 30 35 42 26 5 27 5 29 3 60 34 45 55 6 27 5 30 3 33 5 120 42 61 78 30 35 41 3 180 48 5 74 90 32 39 3 48 240 54 5 82 5 95 34 44 54 5 300 60 88 95 36 48 60 Average Diff 0 7 0 4 0 4 f 0 0 2 1 6 6 2 3 Minimizing Undesirable Factors Other factors may be detrimental to enzyme or biological activity and can reduce the effectiveness of ultrasonic processing Minimize undesirable factors as fol lo
28. atment time will break down particle size to well under micron and gives a fine homogenate Fossils low power will clean debris from delicate fossils without injury Micro fossils such as pollen can be separated from rocks to help identify the geological age of the strata Removal of rock matrix Gamma globulin the Sonifier Cell Disruptor was used to solubilize protein as one of the steps in the biosynthesis of gamma globulin from rabbit spleen Gangliosides immunochemical and structure studies used ultrasonic treatment as one step Gastric mucosa placing scrapings into a test tube and test tube into new water filled cup horn caps permits these cells to be separated and not broken Graphite molybdenum disulfide an excellent dispersion of this lubricant was made in a silicate binder Guanine produces colloidal suspension in 1 minute Gymnodinium solution completely disrupts in 6 minutes Haemophilus pertussis preparation of successful immunological com plexes Heart muscle 1 gm disintegrates in 6 minutes HeLa Cells disruption to free virus in a few seconds without injury Hemphilus pertussis an immunological compound prepared 100 413 016 Rev C A 5 Appendix A Applications BRANSON Herpes virus may be quickly released without injury Histoplasma capsulatum ultrasonic treatment for 7 minutes completely rup tured cells prepared by formalin fixation Good enzyme activity was obtained Human serum proteins u
29. c treatment will produce complete disruption Sulfapyridine excellent dispersion in less than 1 minute Continued ultrasonic treatment will produce complete disruption Synovial fluid ultrasonic treatment is an excellent means ofreducing fluid vis cosity It is simpler and faster than using hyaluronidase T pyriformis excellent disruption 8 enzymes released Tablets complete disruption in 2 40 seconds depending on type Tea excellent extraction Tetrahymena disrupts in a few seconds Enzymes which have been monitored include succinate lactate B hydroxy butyrate glutamate and DPNH oxidases DPNH cytochrome C reductase and ribonuclease Activity of DPNH oxidase was twice that of the best previous experiments Thermoactinomyces disruption of hyphae Homogenization of protein com plex without denaturation Thermophile negative good disruption within 2 minutes Thermophilic bacillus Isocitrate lyase was extracted from a spore forming bacillus similar to Stearothermophilus A washed cell paste suspended in a phos phate buffer was sonified 1 2 minutes and the supernatent was used for enzyme experiments without further treatment Extracts could be frozen and stored with out loss of activity Thiouric acid dissolved in a few seconds Thymus cells complete disruption in 15 seconds Tissue culture cells complete disruption in a few seconds To avoid damage to free organelles and to obtain intact Lyososomes use low power a
30. cides ultrasonic treatment resulted in a 16 fold improvement in the potency ofthe antigen used with Micro crystalline Cellulose as a thin layer adsor bant for chromatographic separation Phosphatidate phosphohydrolase the most potent inhibitors for this enzyme were obtained by making five dispersions with the Sonifier Cell Disrup tor Phospholipid micelles produced stable preparations for an indefinite period Plant cells 30 packed plant cells W V and distilled water depending on type can be completely disrupted in 1 15 minutes Plant tissue 1 gm dried tissue suspended in alcohol disintegrates in about 5 minutes Platelets complete disruption according to size from 20 seconds to 4 minutes Pneumococei preserved in formalin for several years completely disrupts in 6 minutes Polio virus excellent disruption of this most difficult virus Powders broken down to a small relatively uniform particle size PPLO complete disruption in 2 minutes Propionobacteria releases sulfhydro groups intact 70 ml of 20 suspension processed for 10 minutes Propionobacteriom shermanii 2 minutes for extraction of citrate synthose Proteus excellent disruption Pseudomonas aeruginosa rapid complete disintegration Pseudomonas fluorescens 2 gm wet wt in 10 ml completely disrupts in 1 minute A 8 100 413 016 Rev C 250 450 Sonifier Appendix A Applications Instruction Manual Pulmonary cytodiagnosis the mucous in sputum
31. ction Manual Appendix D Accessories Parts List Appendix D Accessories Parts List D 1 Accessories Parts List D 1 D 1 Accessories Parts List Table D 1 Accessories Parts List BRANSON ITEM DESCRIPTION EDP No Disruptor 1 2 dia stepped w grad 101 147 036 Horns scale 1 2 dia stepped tapped 101 147 037 1 2 dia stepped solid 101 147 038 3 8 dia stepped solid 101 147 039 1 2 dia exponential tapped 101 147 040 1 2 dia exponential solid 101 147 041 1 2 dia catenoidal solid 101 147 042 3 4 dia stepped solid 101 147 043 1 dia threaded body solid 101 147 044 3 4 dia solid high gain 101 147 035 100 413 016 Rev C D 1 Appendix D Accessories Parts List BRANSON Table D 1 Accessories Parts List BRANSON ITEM DESCRIPTION EDP No Cup Horn Permit materials to be treated while isolated in small test tubes Cups have transparent plastic bodies 2 diameter 101 147 047 3 diameter 101 147 048 High intensity with flow thru ports 101 147 046 with water jacket Continuous Permits continuous processing of 100 146 171 Flow Attach low viscosity materials with rates ment up to 38 liters hour Designed pri marily for emulsifying dispersing and homogenizing this attachment will disrupt most cells with the exception of the more difficult types Materials being treated may be passed through the attachment
32. d Mycoplasma antibody a suspension of Campo W cells treated for 5 minutes gave 12 lines with the sera in a gel diffusion test The extract was estimated to con tain 12 75 mg protein per ml by Blaret reaction Myeloma tumor cells complete disruption in 10 minutes 30 disruption in 2 minutes Myleran made colloidal suspension and dissolved in approximately 1 minute N crassa nucleus was isolated and purified from conidial extracts after 5 min utes treatment Naeglerigruberi this free living soil amoeba was treated ultrasonically to release sub cellular infectious material Neurospora 40 ml 4 minutes produced more protein than freeze thawing for study of enzymatic synthesis of cystathionine Nocardia ostenodes breaks clumps and disintegrates in less than 10 minutes Nucleoprotein extracted from tissue May be degraded selectively 100 413 016 Rev C A 7 Appendix A Applications BRANSON Oil and water emulsions permanent stable emulsions in a few seconds Par ticle size reduced to less than micron each case slightly different Oil in water water in oil phases can be obtained in same vessel Oyster shell small clean hole can be drilled with micro tip in 3 minutes No cracking is produced Paracolon excellent disruption Parasites separated from red cells in a few seconds Pasteurella pestis complete disruption in 30 minutes using high power Penicillium complete disruption in 3 minutes Pesti
33. e 1s activated Adapter board has failed Harmonics module has failed CE units only Full scale or high meter deflection horn unloaded N Re Power supply module has failed Horn or tip have failed Power supply operates continuously but does not operate in PULSED mode Pulse mode timer or circuitry failed A slight electric shock is felt when touching the Sonifier unit NR Unit is improperly grounded Line cord has failed 100 413 016 Rev C 250 450 Sonifier Appendix A Applications Instruction Manual Appendix A Applications Actinomyces 3 minutes of sonifying produced excellent disruption with 50 protein released and excellent enzyme activity Actinomycin D suspended in 3 minutes Aerobacter aerogenes excellent breakage with better enzyme release than any other method A low power setting can release sulfatase activity into the supernate with no obvious disruption of the majority of cells Aerobacter suboxydans excellent breakage but requires higher power than A aerogenes Algae secendesmus 10 ml concentrated solution completely disrupts in 1 minute Alkaloids total amount and speed of extraction are greater using the Sonifier Cell Disruptor than with standard methods Extraction from ipecac root in 30 sec onds vielded more alkaloid than Soxhlet extraction in 5 hours Antibioticus monocellular elements from surface grown colonies obtained in 1
34. eed to change a horn or converter use the spanner wrench provided NEVER attempt to remove a horn by holding the converter housing in a vise If necessary secure the largest portion of the horn in a soft jawed vise When con necting a horn to the converter use the following procedure 1 Clean contacting surfaces of converter and horn and remove all foreign matter from threaded stud and threaded hole 2 Use the Mylar washer provided in between the converter and required probe Do not apply grease to the threaded stud 4 5 2 Connecting Tip to Horn Figure 4 2 Methods for Connecting Removing Horn from Converter 100 413 016 Rev C 4 5 Chapter 4 Installation and Setup Connecting and Removing Tips Horns and BRANSON Converters A standard flat tip recommended for processing liquids is supplied with tapped horns Other tip configurations are available for experimental work on applica tions where the ultrasonics vibrations are transmitted directly into a solid work piece The shape of the horn influences the direction in which the ultrasonic vibrations are delivered from the horn You will find a list of available tips in Appendix D Accessories Attach tips to horns as follows 1 Clean contacting surfaces of horn and tip and remove any foreign matter from the threaded stud The tip must be installed clean and dry or the power supply may not tune and operate correctly 2 Hand assemble tip to horn 3 Usin
35. egration of difficult tissues Stainless steel construction Cell bot tom holds up to 6g of tissue A water jacket is provided for cooling Reduces mechanical noise generated during liquid pro cessing to a normal level Especially useful when using a cell disruptor for extended periods 25 to 28 db cavita tion noise reduction 101 021 007 101 063 275 Glass Beads For cell homogenization Waterproof 1 pound packages 25 micron diameter 201 002 003 1 2 Wave Hom Extends 1 2 dia disruptor horn an additional 1 2 wave length apprx 5 at a 1 2 diameter 101 147 049 100 413 016 Rev C Appendix D Accessories Parts List BRANSON D 6 100 413 016 Rev C
36. energy 0 to 15 minutes or to HOLD Use HOLD in either the PULSED or CONTINUOUS Modes if you require an undetermined exposure time and ultrasonic energy will be on indefinitely Set the timer anywhere from 0 to 15 minutes to either PULSED or CONTINUOUS Modes and ultra sonic energy will switch off at the end of the pre determined period 2 Duty Cycle dial In the PULSED Mode the ultrasonics are pulsed at a fixed repetition rate of one pulse per second The Duty Cycle control varies the duration of the ultrasonic pulse For example if you choose a 10 setting ultrasonics will be on for 10 of every second if you choose a 90 setting ultra sonics will be on for 90 of every second 2 4 100 413 016 Rev C 250 450 Sonifier Chapter 2 Introduction User s Manual 250 450 Sonifier Power Supply Front Panel Table 2 1 Front Panel Device and Function Item Function 3 ON OFF Applies main power to the unit Lights when Switch power is on 4 Loading Meter Indicates level of ultrasonic power delivered to your sample 5 Output control Controls the amplitude i e peak to peak motion of the ultrasonic vibrations Clockwise rotation increases amplitude For further information refer to Table 6 2 Horn Amplitudes When using micro tips maximum output control setting is 6 100 413 016 Rev C 2 5 Chapter 2 Introduction Model 250 450 Sonifier Power Supply Rear BRANSON Panel
37. er Output Loss 7 2 7 3 Preventive Maintenance 7 3 7 4 Reconditioning the Stack Interface 7 3 7 4 1 Refacing the Mating Surfaces 7 4 7 4 2 Stud Replacement 7 6 7 4 3 Horn Tip Cleaning 7 7 7 4 4 System Trouble Analysis Chart 7 7 100 413 016 Rev C 7 1 General Information BRANSON 7 1 General Information If maintenance is required contact Branson Product Support See Section 1 6 How to Contact Branson for contact information If you have a problem operating your unit 1 Refer to Table 7 1 to locate the symptom that most clearly describes your problem 2 See Appendix B Parts List for available replacement parts 3 Refer to Chapter 1 Safety and Support for equipment return infor mation 7 2 Maintenance and Troubleshooting The Digital Sonifier is a self contained system that requires no internal servicing except for a protective fuse and there are no user serviceable parts inside the unit The ultrasonic tooling horns and tips may require periodic inspection and main tenance to ensure optimum performance The tooling components are subject to wear and may require replacement after a period of time depending on the appli cations If you have a problem operating your unit refer to Table 7 1 Symptom Probable
38. er supply If it senses conditions that could cause the power supply to overload it will shut the power module down To reset the power supply after an overload condition has been sensed turn the power switch off and then turn it back on Autotune optimizes the operating frequency of the power supply so that it is consistent with the acoustic tooling To facilitate the auto tune process the power supply performs a seek function upon first powering up the unit When first powering up the unit will perform a brief cycle at low amplitude so that the power supply can determine information about the acoustic tooling including it s operating frequency The output meter on all Analog Sonifiers now reports Watts It is no longer necessary to use the tables in the manual to determine output watts based on meter reading vs output control setting 100 413 016 Rev C 2 3 Chapter 2 Introduction 250 450 Sonifier Power Supply Front Panel BRANSON 2 3 250 450 Sonifier Power Supply Front Panel Figure 2 2 Power Supply Front Panel 7 N 7 a b Both illustrations represent a Sonifier Power Supply Front Panel with the exception that the image shown as b is intended for use in Japan Table 2 1 Front Panel Device and Function Item Function 1 Timer Times the application of ultrasonic
39. ermophillic spore form 98 disruption of 70 ml of 40 suspension in 15 minutes Bacillus brevis 1 15 W V in 3 minutes Bacteroides Symbiosis 1 phosphofructokinase a soluble enzyme has been isolated from this anaerobe by ultrasonic treatment A 25 ml suspension was son ified for 10 minutes and centrifuged at 36 000 xg for 10 minutes Baker s yeast saccharomyces cerevisiae see Yeast Blastomyces dermatitidis 95 disruption in 3 minutes Blood cells red and white cells can be disrupted in a few seconds Boll weevil tissue complete homogenization in a few seconds Bone compact bone can be sonified and processed for microscopic sections in minutes Other methods can reguire up to a week Bone specimens treated in this way vielded large numbers of intact cells with little distortion Malignant criteria were easily recognized Tumor types studied were osteosarcoma chondrosar coma liposarcoma chordoma metastic bronchogenic sguamous and benign giant Bone can be decalcified without injury to the cells processed for micro scopic sections and diagnosed in a short time Other methods reguire extensive treatment time Brain stem and adrenal gland ultrasonic treatment dispersed 10 mg sam ples in 10 ml fluid which is usually difficult without substantial loss of material The suspension was analyzed for nucleotides Brain tissue disintegrates instantly A 2 100 413 016 Rev C 250 450 Sonifier Appendix A Applications Instr
40. g spanner wrench on horn and open end wrench on tip tighten tip See Figure 4 3 Torque specifications for the various threaded tips follow 1 4 20 thread tighten at 90 inch Ibs 10 16 Newton meters 1 4 28 thread tighten at 110 inch lbs 12 42 Newton meters 3 8 24 thread tighten at 180 inch Ibs 20 33 Newton meters Figure 4 3 Connecting or Removing Tip from Horn 4 6 100 413 016 Rev C 250 450 Sonifier Chapter 4 Installation and Setup User s Manual Guards and Safety Equipment 4 6 Guards and Safety Equipment Although the Sonifier operates outside the normal range of human hearing some applications can create audible noise above 85dB If an uncomfortable level of noise is present the operator should wear ear protection for safe operation Appropriate eye protection should be worn when operating the Sonifier to pre vent possible splash injury originating in the solution The Ultrasonic Horn can cause injury and or equipment damage during operation To avoid injury or accident never touch the Ultrasonic Horn while the System is turned on and do not allow the Horn to come in con tact with solid vessels or supports 100 413 016 Rev C 4 7 Guards and Safety Equipment BRANSON 4 8 100 413 016 Rev C 250 450 Sonifier Chapter 5 Technical Specifications User s Manual Chapter 5 Technical Specifications 5 1 Specifications D 2 5 1 1 Electrical Specifications
41. he Stack Interface BRANSON 7 4 1 Refacing the Mating Surfaces Never clean the Converter or Horn mating surfaces with a buffing wheel 1 Disassemble the Converter Horn Stack and wipe the mating sur faces with a clean cloth or paper towel 2 Examine all mating surfaces If any mating surface shows corro sion or a hard dark deposit recondition it 3 Examine the knurled end of the stud If worn replace the stud Threaded studs cannot be reused in titanium horns If you remove the stud from the horn you must replace it with a new stud 4 Tape a clean sheet of 400 or finer grit emery cloth to a clean smooth flat surface such as a sheet of plate glass 400 Emery Cloth Be careful to avoid tilting the part and losing flatness of the surface Doing so may make the system inoperative due to improper mating sur faces 7 4 100 413 016 Rev C 250 450 Sonifier Chapter 7 Maintenance User s Manual Reconditioning the Stack Interface Lapping Procedure 5 Holding the part to recondition place the interface surface on the emery cloth Grasp the part at the lower end with your thumb over the spanner wrench hole and lap the part in a straight line across the emery cloth Do not apply downward pressure The weight of the part alone provides sufficient pressure Rotate the part 120 degrees 1 3 to the next hole Stroke the part an equal number of times at each rotation 2 or 3 Pick up the part and lap it
42. he tip and threads which must be thoroughly dry and clean prior to reassembly Using a spanner wrench on the horn and an open end wrench on the tip install the tip tightly using the following torque specifications e 1 4 20 tighten at 90 inch Ibs 10 16 Newton meters e 1 4 28 tighten at 110 inch Ibs 12 42 Newton meters 3 8 24 tighten at 180 inch Ibs 20 33 Newton meters The 250 450 Sonifier does not require retuning 7 3 Preventive Maintenance Periodically Clean the Equipment Periodically disconnect the unit from power remove the cover and vacuum out any accumulated dust and debris Remove material adhering to the circuit boards cooling intake vents and exhaust ports External covers may be cleaned with a damp sponge or cloth using a solution of mild soap and water Do not allow clean ing solution to enter the unit 7 4 Reconditioning the Stack Interface Ultrasonic system components work most efficiently when the mating surfaces of the Converter and Horn combination also called a Stack are flat in solid con tact and free from fretting corrosion Fretting corrosion refers to a black crusty build up resulting from friction between metal parts that appears on the Stack mating surfaces Poor contact between mating surfaces wastes power output makes tuning difficult increases noise and heat and may cause damage to the converter 100 413 016 Rev C 7 3 Chapter 7 Maintenance Reconditioning t
43. hen you contact the Product Support department to help identify the problem Before calling determine the following information 1 Your company name and location 2 Your return telephone number 3 Have your manual with you If troubleshooting a problem refer to Chapter 7 4 Know your equipment model and serial numbers found on a data label on the units Information about the Horn part number gain etc or other tooling may be etched into the tooling 5 What horn and accessories are being used 6 What are the setup parameters and mode 7 Describe the problem provide as much detail as possible For example is the problem intermittent How often does it occur How long before it occurs if you are just powering up 1 6 100 413 016 Rev C 250 450 Sonifier Chapter 1 Safety and Support User s Manual Returning Equipment for Repair 8 Listthe steps you have already taken 9 What is your application including the materials being processed 10 Have a list of service or spare parts you have on hand tips horns etc Notes 1 7 Returning Equipment for Repair To return eguipment to Branson you must first obtain an RGA number from Branson or the shipment may be delayed or refused Call the Repair department to obtain a Returned Goods Authorization RGA number If reguested the Repair department can send you a facsimile of the Returned Goods Authorization form to fill out and return with your egu
44. ime of longer than 15 minutes is required Set Duty Cycle Control to the required pulse duration The lower the number setting the shorter the pulse and the slower the resulting temperature rise of the solution Switch power On and adjust Output Control to the required intensity When using micro tips do not advance the output control beyond 6 or permanent damage to the micro tips can result Do not allow micro tips to touch anything other than the solution The stress of touching any container can cause fracture of the tip or container 6 2 100 413 016 Rev C 250 450 Sonifier Chapter 6 Operation User s Manual Operating Considerations Prior to processing an unknown sample estimate variables such as pro cessing time output control setting starting temperature and maximum permissible temperature of sample Remember that the temperature of a liquid will rise from the effect of ultrasonically induced cavitation First process a small sample ofthe specimen using your estimated con trol variables When unsure initial test should be conducted with short time and low amplitude setting Examine the results against your objective Several trial runs may be necessary Based on results obtained adjust the variables until you obtain satisfactory results experimenting with one variable at a time 6 2 Operating Considerations The following sections discuss operating techniques under varying conditions 6 2 1 Limiting Temperatu
45. ion and homogenization E coli 2 gm wet wt in 10 ml solution completely disrupts in 40 seconds The Sonifier Cell Disruptor has been used extensively in research on this organism Egg whites can bereduced to ahomogeneous pipettable solution in 15 seconds on low power Ehrlich ascites disrupt in a few seconds Electron microscopy apertures are quickly cleaned Emulsions 10 ml of most light mixtures become semi permanent emulsions in about 1 minute without emulsifiers Average particle size is usually well under 1 micron Sterile emulsions can be prepared by ultrasonic treatment for feeding to germ free animals A 4 100 413 016 Rev C 250 450 Sonifier Appendix A Applications Instruction Manual Enterococcus excellent disruption Erwina cartovara complete disruption in 1 2 minutes depending on cell con centration Erythrocytes disrupts in a few seconds Euglena gracilis completely disrupts in a few seconds to isolate chloroplasts Eugoena complete disruption in 12 minutes 90 disruption in 8 minutes with pigment released Fat extraction fatcan be emulsified without injuring tissue with proper power selection Lipid layer can be stripped from spores and mycobacteria Fibrin complete suspension of 125 gm in 30 minutes Fish gill complete disruption of 20 mg in 30 seconds Fish tissue tissue homogenization for extractions and excellent particle size reduction in 8 minutes per 10 gm Fluorocarbons extended tre
46. ipment Branson Repair Department C O Zuniga Logistics TD 12013 Sara Road Killam Industrial Park Laredo Texas 78045 U S A direct telephone number 877 330 0405 fax number 877 330 0404 Provide as much information as possible that will help identify the need for repair Include a copy of page 1 8 with your information filled in Carefully pack the equipment in original packing cartons e Clearly label all shipping cartons with the RGA number on the outside of cartons as well as on your packing slip along with the reason for return e Return general repairs by any convenient method Send priority repairs by air freight You must prepay the transportation charges FOB Laredo Texas U S A 100 413 016 Rev C 1 7 Returning Equipment for Repair BRANSON Make a copy of this page and return it with the product Before sending equipment for repair provide as much information with the equip ment to help determine the problem with the system Fill in any details below or on a separate sheet 1 Describe the problem provide as much detail as possible For exam ple is this a new problem Is the problem intermittent How often does it occur How long before it occurs if you are just powering up 2 What are the setup parameters 3 What is your application e g continuous pulse temperature etc 4 Name and phone number of the person most familiar with the problem
47. ips are designed to give maximum mechanical energy output Since they operate close to the stress limits oftitanium Branson cannot guarantee Microtips against failure CONTACT YOUR BRANSON REPRESENTATIVE OR BRANSON ULTRA SONICS CORPORATION DANBURY CONNECTICUT SHOULD YOU HAVE ANY QUESTIONS CONCERNING HORN QUALIFICATION 1 800 732 9262 100 413 016 Rev C 1 5 How to Contact Branson BRANSON 1 6 How to Contact Branson Branson is here to help you We appreciate your business and are interested in helping you successfully use our products To contact Branson for help use the following telephone numbers or contact the field office nearest you e Customer Service 800 732 9262 Technical Support 800 732 9262 Applications Assistance 203 796 0532 Parts Store 877 330 0406 Central Time Zone Repair department 877 330 0405 Central Time Zone Danbury Main Number all Departments 203 796 0400 Eastern Time Zone Tell the operator which product you have and which person or department you need If after hours please leave a voice message with your name and return tele phone number 1 6 1 Before Calling Branson for Assistance This manual provides information for troubleshooting and resolving problems that could occur with the equipment see Chapter 7 If you still require assistance Branson Product Support is here to help you The following questionnaire lists the common questions you will be asked w
48. lls and tissues adding glass powders 5 microns to 0 5 mm will materially decrease treatment tissues especially when used in conjunction with the standard high intensity micro tip A ratio of part glass powder to 2 parts liquid is recommended 100 413 016 Rev C 6 7 ak a AEE BRANSON 6 3 Horn and Micro Tip Amplitudes Table 6 2 Horn Amplitudes Output Control Setting 1 5 10 max Part No Description 101 147 037 1 2 Dia Tapped Stepped Disruptor 21 0 76 0 145 0 0008 003 0057 101 147 042 1 2 Dia Solid Catenoidal 21 0 74 0 145 0 0008 0029 0051 101 147 041 1 2 Dia Solid Exponential 10 0 34 0 65 0 0004 0013 0025 101 147 039 3 8 Dia Solid Stepped Disruptor 36 0 125 0 240 0 0014 0049 0094 100 147 043 3 4 Dia Solid Stepped Disruptor 9 5 33 0 63 0 0004 0013 0025 101 147 035 3 4 Dia Solid High Grain 19 0 68 5 130 0 007 1 0027 0051 101 147 044 1 0 Dia Solid Stepped 6 3 21 5 40 5 0002 1 0008 0016 All measurements in Microns 6 8 100 413 016 Rev C 250 450 Sonifier User s Manual Chapter 7 Maintenance Chapter 7 Maintenance 7 1 General Information 7 2 7 2 Maintenance and Troubleshooting 1 2 7 2 1 Tip Erosion 7 2 7 2 2 Pow
49. ltrasonic treatment causes a reproducible change in the electrophoretic behavior of normal human serum consisting of an increase in material migrating in the x and b globulin zones with a reduction in the albumin and y globulin fractions Hydrocortisone smaller crystals were produced by ultrasonic treatment Hydrophilic vegetable gums disperses and solubilizes hydrophilic vegeta ble gums in water makes dispersions of added particulate matter Intracellular membrane disruption and particle size reduction obtained in 30 60 seconds Isoenzymes selectively activated with respect to time and intensity of treat ment Kidney 1 gm disintegrates in 3 minutes Kidney stones easily broken in seconds in vitro Klebsiella excellent disruption L arabinosis complete disruption to free virus in 2 minutes without injury Lactobacillus 0 5 gm in 15 ml completely disrupts in 11 minutes Excellent release of acetokinase Lenconostoc mesenteroides ultrasonic treatment for 15 minutes using high power for disruption Leukocyte Iysozyme activity in myelocytic leukemia the cell suspen sion was ultrasonically treated and samples assayed for lysozyme activity The lysozyme concentration of the leukocytes ug 10 cells was determined Linoleic acid made suspension in water in 30 seconds Liver tissue 1 gm homogenizes in less than 1 minute Lung tissue 1 gm homogenizes in 2 minutes Lymphacytis complete disruption in 15 seconds Ly
50. m However it is faster easier and equally effective to sterilize horns by immersion in a beaker of alcohol or other disinfectant and then turning the power on for a few seconds This technique also cleans or removes unwanted residue on the horn or tip 6 2 5 Disrupting Tissues and Solids You can effectively homogenize or disrupt many kinds of tissue and other solids Energy radiates only from the horn s tip The energy is most concentrated within 1 2 inch 13 mm of the face of the tip When you treat tissue or solids already in solution the freely moving cells or particles pass before the face of the tip many times during the process When you treat a solid piece however the energy pat tern from the tip of the horn has a tendency to push or repel the solid away from the tip The solid does not receive treatment but simply spins or circulates around the container 6 6 100 413 016 Rev C 250 450 Sonifier Chapter 6 Operation User s Manual Operating Considerations You can effectively treat all but the most difficult of materials by the following two steps 1 Homogenize the tissues or solids by placing them in a high speed blender with the solution 2 Insert the horn in the liquid sample for complete disruption To disrupt solid pieces especially those that are extremely resistant to breakage place the horn directly over the tissue or right against it 6 2 6 Using Glass Powders with Solution To disrupt difficult ce
51. mate rial rock surfaces between polishing stages S faecalis excellent disruption in 1 minute S fragilis 5 minutes yielded excellent release of galactokinase more than any other method Subcellular particles may be extracted or disrupted Saliva glands complete disruption Salmonella various culture media or phosphate buffered saline disintegrated between 40 and 50 in 10 20 minutes Sonifying was one step in an improved assay for enzyme thiogalactosize transacetylase 100 413 016 Rev C A 9 Appendix A Applications BRANSON Salmonella typhimurium and enteritidis bacteria were suspended in 1 300 volume of original culture sonified for 4 minutes and centrifuged for 20 min utes at 20 000 9 Extracts were found to catalyze the synthesis of cytidine diphos phate 3 6 dideozyhexoses Schistosoma mansoni complete disruption Sedimentary rock completely disperses flocs with the release of all bound silt and clay particles Sediments ultrasonic treatment disperses fine material permitting quick neat separation of sand from silt and clay fractions Serial number restoration used in crime laboratories to restore obliterated serial numbers Serratia marcescens complete breakdown of a concentrated solution in 1 minute Serum quickly homogenized Serum cholinesterase activated by ultrasonic treatment Different cholinest erase isoenzymes may be activated selectively and inactivated selectively Shale excellent
52. more than once to obtain the desired results A water jacket input output and overlow connections are pro vided For use with horns having outside threads Sealed Atmo Used for batch treatment of infectious materials Input sphere Treat and output connections allow filling and emptying with ment Chamber breaking the airtight seal and permit processing with an inert gas Stainless steel 3 10ml 101 021 001 6 15ml 101 021 002 20 50ml 101 021 003 100 413 016 Rev C 250 450 Sonifier Instruction Manual Appendix D Accessories Parts List Table D 1 Accessories Parts List ITEM DESCRIPTION pr a Sealed Atmo Same as above 101 021 001 series but with cooling sphere Treat water jacket ment Chamber 3 10ml 101 021 004 6 15ml 101 021 005 20 50ml 101 021 006 Wrenches For use with Branson Cell Disruptors Spanner 201 118 019 Open end for 1 2 horn 201 118 010 Flat Tip Replacement for 1 2 horn 101 148 013 Tapered Micro For processing small volumes Attaches to Tips standard tapped disruptor horn Tip amplitude is 3 1 2 times greater than that of standard horn Recommended for difficult applications such as spores fungi yeast muscle and con nective tissue Excellent results on volumes ranging from 3 20ml in a comparatively short time 1 8 diameter 101 148 062 3 16 diameter 101 148 069 1 4 diameter 101 148 070 100 413 01
53. mphocyte nuclei complete disruption in 6 minutes Lymph gland direct injection lymphography with a modified radiopaque emulsion was obtained by ultrasonic treatment in a functional procedure produc ing lymphatic structure detail A 6 100 413 016 Rev C 250 450 Sonifier Appendix A Applications Instruction Manual Lysossomes released enzymes quickly Malaria prolozoa fast excellent disruption Maple bark spores complete disruption in 14 minutes Measles disruption of virus antigen clumps present in infected cells on low power Ultrasonic treatment increased antigen titer 4 8 fold Methanobacillus omelianskii 1 gm cells wet wt ml completely disinte grates in 2 minutes for assaying methane Microbacterium lacticum ultrasonic treatment used for malate dehydroge nase extraction Micrococci a 13 ml solution completely disrupts in 15 minutes Micrococcus lactiliticus 75 ml of a 20 suspension was disintegrated in 15 minutes and a good yield of the enzyme Xanthine dehydrogenase extracted Mineral rock excellent for cleaning surfaces between polishing stages Mitochondria separates from cells without injury Mitochondria themselves can be broken with longer ultrasonic treatment Inner membrane sub units also isolated Muscle tissue 1 gm homogenized in 4 minutes heart muscle in 6 minutes Mycobacteria a 20 ml growing media completely disrupts in 14 minutes Clumps break quickly An immunological compound prepare
54. o per sonal injury 100 413 016 Rev C 1 3 intended Use of the System BRANSON 1 3 Intended Use of the System The 250 450 Sonifiers can be used to disrupt cells bacteria spores or tissue and are ideal for initiating and accelerating chemical biochemical and physical reac tions and degassing liquids With the 250 450 Sonifiers you can prepare an emul sion to 01 micron homogenize immiscible liquids polymerize some materials and de polymerize others Appendix A lists the effects of irradiating a variety of biological materials with ultrasonic energy at 20 kHz The 250 and 450 Sonifiers differ in their output power ratings with the output con trol at maximum setting The 250 maximum available output power is 200 watts the 450 maximum available output power is 400 watts If the Sonifier is used in a manner not specified by Branson the protec tion provided by the equipment may be impaired 1 4 Regulatory Compliance Sonifier sold in Europe after 1 1 01 are CE compliant They have new part num bers They conform to EN55011 EN 61010 1 EN61000 3 2 EN61000 3 3 and EN61326 They take advantage of Branson latest System Protection and Autotune technology 1 4 100 413 016 Rev C 250 450 Sonifier Chapter 1 Safety and Support User s Manual Warranty 1 5 Warranty Refer to the Terms and Conditions of Sale found on the back of your Invoice for information about the product Warranty issued of your Branson products
55. ons 5 1 Specifications 5 2 5 1 1 Electrical Specifications 5 2 5 1 2 Mechanical Specifications 5 3 5 1 3 Dimensions 5 4 Chapter 6 Operation 6 1 General Operation 6 2 6 1 1 Timed Operation 6 2 6 1 2 Continuous Operation 6 2 6 1 3 Pulsed Operation 6 2 6 2 Operating Considerations 6 3 6 2 1 Limiting Temperature Rise 6 3 6 2 2 Vessel Capacity and Speed of Temperature Rise 6 5 6 2 3 Minimizing Undesirable Factors 6 5 6 2 4 Sterilizing and Preventing Cross Contamination 6 6 6 2 5 Disrupting Tissues and Solids 6 6 6 2 6 Using Glass Powders with Solution 6 7 6 3 Horn and Micro Tip Amplitudes 6 8 Chapter 7 Maintenance 7 1 General Information 7 2 7 2 Maintenance and Troubleshooting 1 2 7 2 1 Tip Erosion 7 2 7 2 2 Power Output Loss 7 2 7 3 Preventive Maintenance 7 3 7 4 Reconditioning the Stack Interface 7 3 7 4 1 Refacing the Mating Surfaces
56. quipment is operating properly proceed as follows a Mount 1 2 disruptor horn with flat tip if tapped to converter b Set Output Control to 5 Timer to HOLD and ON OFF switch to ON c Record meter reading with horn in air Meter Reading Horn Size d Fill 500 ml pyrex beaker to 500 ml level with room temperature tap water e Immerse horn tip halfway to 250 ml mark Set ON OFF switch to ON and record meter reading Meter Reading Perform this test and compare readings periodically to ensure that the equipment is operating satisfactorily These readings are a base for comparison to readings taken later on The above meter reading with horn in air should have no greater than a 10 variation when compared to any future metering readings taken with horn in air Likewise the above meter reading with horn tip immersed halfway to 250mL mark should have no greater than a 10 variation when compared to any future meter readings taken with the horn tip immersed halfway to 250 mL mark If readings are not within tolerance refer to the Trouble Analysis Chart on Chap ter 7 Maintenance 4 4 100 413 016 Rev C 250 450 Sonifier Chapter 4 Installation and Setup User s Manual Connecting and Removing Tips Horns and Convert ers 4 5 Connecting and Removing Tips Horns and Converters 4 5 1 Connecting Horn to Converter The 250 450 Sonifier horns have been torqued onto the converter at the factory Should you n
57. re Rise An important objective in ultrasonic emulsification is to keep processed samples cool Selection of the proper processing vessel and cooling bath resolves most heating problems While any type of vessel can be used to hold the sample the shape of the vessel depends primarily on the volume to be processed For small volumes choose the smallest diameter vessel that allows the probe to be inserted without touching the sides of the vessel This minimized diameter size raise the height of the liquid permitting greater surface area to be exposed to the cooling bath for more effective heat transfer 100 413 016 Rev C 6 3 Operating Considerations BRANSON Based on heat transfer characteristics the following vessel materials are recom mended listed in decreasing order of heat conductivity Best Aluminum e Stainless Steel Thin wall Glass Thick wall Glass Poorest Plastic Immersing the processing vessel in a simple ice water bath 0 C provides suffi cient cooling for larger volumes if required treatment times are short If temper ature rise is too great with this method consider using the following e Ice salt 6 C Ice alcohol 14 C Dry ice alcohol water 30 to 40 C For smaller volumes with less than 30 seconds treatment time an ice water bath is sufficient For longer periods especially when high power is required a lower temperature bath is required 6 4 100 413 016 Rev C 250 450 Sonifier Ch
58. sity Cup Horn has a concave bottom that focuses the ultrasonic energy on the bottom ofthe tube The larger units with diameters of 2 or 3 inches allow for the immersion of multiple test tubes The larger Cup Horns have clear plastic walls which permit easy viewing of the activity in the tubes during processing Both types of Cup C 2 100 413 016 Rev C 250 450 Sonifier Appendix C Accessories Instruction Manual Horns are designed to allow chilled water to circulate through the Cup Horn to prevent heating of the solution as a result of the ultrasonic activity The Cup Horn 1s available with a water cooling device which allows water to flow through the cup keeping the material cool The bottom of the test tubes should not be in contact with the surface of the ultrasonic horn which could cause breakage C 3 Flow thru Horn Designed for pharmaceutical research the flow thru horn has two inputs or ori fices at the non vibrating nodal point of the horn A pre mixed substance is fed through one of the units while the horn 1s ultrasonically activated Because two inputs are available two different types of material can be treated simultaneously for mixing or emulsifying The processed solution exits at the tip of the horn The horn may be used as a continuous flow device to collect the solution in one large vessel Or by using a special switching an instantaneous ON OFF action permits multiple tubes or vials to be filled C 4 Rosette
59. t short expo sure 100 413 016 Rev C A 11 Appendix A Applications BRANSON Toxin and antitoxin one example of many Toxin preparations of whole cell lysate WCL of the Inaba serotype strain 569E of the classic biotype of cholera vibrio were grown on 3 Bacto peptane agar and harvested in distilled water at 18 hours The unwashed suspensions were solubilized ultrasonically clarified by centrifugation and the supernat freeze dried for the titration of cholare toxin in the rabbit ileal loop Toxoplasma gondii can be separated from white blood cells without injury Transplantation antigens were extracted from spleen thymus and lymph nodes Trichomonas foetus Complete disruption in a few seconds Triolein Complete stable emulsion in 2 minutes Trypanosomes concentrated 10 ml solution completely disrupts in 4 minutes Uterus muscle a 1 5 gm 3 cc solution completely disrupts in 3 minutes for coenzyme Q determination Vaccines numerous advantages such as more antigenic material released than usual and the production of vaccines not obtainable by classified methods Various bacilli complete disruption in 3 minutes Vibrio comma excellent disruption Virus extraction excellent for experimental vaccines Evidence of breakage of virus antibody bonds Virus can be extracted without damage at low power or broken at high power Vitamin E 30 seconds of ultrasonic treatment put material in solution with a resultant permanen
60. t suspension W138 virus cell free V 2 virus obtained in 30 seconds using 6 ml of Veronal buffer with W138 cells containing V 2 virus Yeast complete disruption in 3 10 minutes Complete disruption of 9 grams pressed yeast in 18 ml buffer in 8 minutes Protein release 52 mg ml from an aged sample A 12 100 413 016 Rev C 250 450 Sonifier Appendix B Parts List Instruction Manual Appendix B PartsList Table B 1 Replacement Parts Item Description EDP Number Replacement 5 00 Amp for Model 450 117V 200 049 121 Fuses Mylar Washer Kit Mylar Washer 20Khz 100 063 357 100 098 120 Stud Replacement studs for ultrasonic A accessories a 100 098 249 100 098 370 Converter 102C Converter CE compliant 101 135 066 For use with Branson Sonifier Cell Disruptors 201 118 019 Wrenches Spanner Open end for 1 2 disruptor tip 201 118 010 Extends 1 2 diameter disruptor horn te Wave Hor an additional 1 2 wavelength 101 147 049 Extension approx 5 at a 1 2 diameter For more information call Branson 100 413 016 Rev C Appendix B Parts List BRANSON B 2 100 413 016 Rev C 250 450 Sonifier Appendix C Accessories Instruction Manual Appendix C Accessories C 1 Micro Tip C 1 C 2 Cup Horn C 2 C 3 Flow thru Horn C 3 C 4 Rosette Cell C 3 C 5 Continuous
61. uction Manual Brevi bacterium 25 ml disrupts in 20 seconds Brevi bacterium acetylicum approximately 3 minutes to disrupt large sam ples and measure TCA enzyme activity Brine Shrimp complete disintegration in 1 minute Brucella abortes separates easily from leukocytes At least 9 antigens extracted Bull sperm contractile protein is easier to extract from tails after sonifying C butyricum C cylinrosporum C kluyveri vegetative cells easily dis rupted C pasteurianum 3 minutes disruption for hydrogens reducing Ferredoxin with H2 Calcium mouse Ehrlich ascites tumor cells were sonified for 1 minute to deter mine the amount of bound calcium present Cells were labelled with calcium 45 Candida albicans spores 95 disruption of 1 2 gram dry wt ina 15 ml solu tion in 35 minutes Carbon black excellent small particle suspension Caryophanon latum glucosamine muramic acid alanine glutamic acid and lysine were obtained Catecholamine can be extracted from heart muscle Cellumonas biazotea disruption obtained with retention of malate dehydrogenase activity Chicken spermatozoa complete disrupts in 2 minutes Chlorella completely disrupts in 3 minutes Chloroplasts disrupts in a few seconds Cholesterol apparent permanent suspension in 1 minute in water Chromatography prior ultrasonic treatment of adsorbant in any convenient solvent for a few seconds eliminates aggregates and results in a uniform easily
62. ue culture cells HeLa cells and the com plete range of cells which have low to medium resistance to breakage To prevent foaming or aerosoling while processing small guantities with the tapered or stepped micro tips the use of a conical shaped tube such as an eppen dorf tube or a cut down centrifuge tube is recommended When using micro tips a maximum Output Control Setting of 6 must not be exceeded The micro tip will break if driven at higher amplitudes C 2 Cup Horn Cup Horns allow materials to be treated in small eppendorf style tubes or test tubes without immersing the ultrasonic horn or micro tip in the material thereby providing completely sterile conditions The Cup Horn is attached directly to the converter and the assembly is mounted upside down on the lab stand or in our Sound Enclosure with the assembly inserted through the bottom With chilled water in the Cup Horn the tubes are suspended in the Cup with the contents ofthe tubes just below the water level Ultrasonic energy is then conducted from the sur face of the horn through the water and tube walls to the contents of the tube Some energy loss may be experienced when applying ultrasonics in this indirect way and processing may take longer than if the ultrasonic horn were immersed directly in the solution There are three types of Cup Horns a high intensity unit that willaccommodate a single test tube and two larger units for multiple test tubes The high inten
63. ws 100 413 016 Rev C 6 5 Operating Considerations BRANSON 6 2 3 1 Foaming or Aerosoling Always place the horn deep enough below the surface of the liquid to ensure there is no violent motion or agitation on the surface This problem is more critical when processing small volumes for example 0 3 to 5 ml when a conical shaped tube or vial such as a cut down Eppendorf tube is recommended The conical shape of this type of container effectively raises the liquid level without increasing the volume thereby permitting the horn to be inserted more deeply below the lig uid surface level If foaming occurs it can be detected by a change in the sound level and a fluctuating reading on the power meter Little or no energy couples reliably to the solution while aerosoling is oc curring and excessive top layer heating will result Remedy this problem by lowering the probe as deep as possible and placing the output power control to position or 2 for a few seconds Then gradually advance the power output control to the level required 6 2 3 2 Discoloration of the Processed Sample If the tip touches the side of a glass tube or beaker small glass particles will be released which gradually discolor the sample to a greyish color Excessive tip cor rosion can also cause a greying or darkening condition 6 2 4 Sterilizing and Preventing Cross Contamination Sterilize horns and tips by removing them from the converter and autoclaving the

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