250 - 450 Sonifier Analog Cell Disruptor User`s Manual
Contents
1. 3 3 4 Operation 4 1 4 1 General Operation 4 1 4 1 1 Timed Operation 4 1 4 1 2 Continuous Operation 4 2 4 1 3 Pulsed Operation 4 2 4 2 Operating Considerations 4 3 4 2 1 Limiting Temperature Rise 4 3 100 413 016 Rev i BRANSON 4 2 2 Vessel Capacity and Speed of Temperature Rise 4 4 4 2 3 Minimizing Undesirable Factors 4 5 4 2 4 Sterilizing and Preventing Cross Contamination 4 6 4 2 5 Disrupting Tissues and Solids 4 6 4 2 6 Using Glass Powders with Solution 4 7 4 3 Testing the Equipment 4 7 4 4 Connecting and Removing Tips Converters 4 8 4 4 1 Connecting Horn to Converter 4 8 4 4 2 Connecting Horn 4 8 4 5 Horn and Micro Amplitudes 4 10 5 General Information 5 1 5 1 General Information 5 1 5 1 1 System Trouble Analysis Chart 5 1 6 Main Electrical Assembly 6 1 250 450 Sonifier Assembly 6 2 6 2 250 Power Supply Parts List2. 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 reguired probe Do not apply grease to the threaded stud 4 4 2 Connecting Tip to Horn Figure 4 1 Methods For Connecting Removing Horn From Converter 4 8 100 413 016 Rev 250 450 Sonifier Chapter 4 Operation User s Manual Connecting and Removing Tips Horns and Convert ers 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 Using spanner wrench on horn and open end wrench on tip tighten tip See Figure 10 Torque specifications for the various threaded tips follow f 1 4 20 thread tighten at 90 inch Ibs 10 16 Newton meters 1 4 28 thread tighten at 110 inch Ibs 12 42 New
3. 2 SYMBOL INDICATES GROUND CONNECTION TO GROUND PLATE MEMORY_STORE MEMORY CLEAR EXT SEEK EXT SEEK 100 413 016 Rev A 6 5 250 450 Sonifier The Effects of Ultrasonic Instruction Manual Irradiation on Various Biological Materials Appendix A_ The Effects of Ultrasonic Irradiation on Various Biological Materials 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 yielded more alkaloid than Soxhlet extraction in 5 hours Antibioticus monocellular elements from surface grown colonies obtained in 1 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
4. 6 3 6 2 1 Domestic 100 132 1622 6 3 6 3 450 Power Supply Parts List 6 4 6 3 1 Domestic 100 132 1623 6 4 6 4 250 450 Interconnect Schematic 6 5 A The Effects of Ultrasonic Irradiation on Various Biological Materials 1 How to obtain Further Assistance B 1 Customer Service Technical Support and Applications Assistance B 1 Equipment Failure Support 1 Returning Eguipment 1 100 413 016 Rev 250 450 Sonifier User s Manual Accessories 54a C4 Micro Tip 1 Horn 2 Flow thru Horn 3 Rosette Cell C 3 Continuous Flow Attachment 4 Soundproof 4 D Accessories Parts List D 1 Accessories Parts List D 1 100 413 016 Rev B iii BRANSON List of Figures 1 1 2 1 2 2 2 3 2 4 3 1 4 1 4 2 6 1 C 1 Model 250 450 Sonifier Main Components 1 3 Supply Front Panel 2 1 Power Supply Rear Panel
5. Connector 4 RF Cable Connects the power supply to the converter 100 413 016 Rev B 2 3 Chapter 2 Introduction Specifications BRANSON 2 3 Specifications 2 3 1 Electrical Specifications Table 2 3 Electrical specifications Controls amp Displays ON OFF Switch Output control Duty Cycle Connector Horn Frequency Line Voltage Power Minimum acceptable output power Minimum Starting Power Temperature Tuning Output control at maximum setting Timer 0 15 Minutes and Hold Output control Duty Cycle Pilot Light Adjusts amplitude of power supply output volt age Range 10 100 of nominal converter ampli tude Intermittent pulse duration adjustable 10 9096 1 pulse per second Continuous Processing Time RF connects 20kHz high voltage to converter connections Model 102C 19 850 20 050 kHz 100V 10V 50 60 Hz 115V 10V 50 60 Hz 200 250 selectable 50 60 Hz Maximum ground leakage current 115V line to ground 1 2 Model 250 150 Watts Model 450 350 Watts Model 250 120 Watts Model 450 300 Watts Operating 41 F 5 C 122 F 40 C Ambient Storage and Shipping 22 F 30 C to 158 F 70 Factory set no tuning necessary 100 413 016 Rev B Chapter 2 Introduction 250 450 Sonifier Specifications User s Manual 2 3 2 Mechanical Specifications Mechanical specifications Table 2 4 Converter
6. 2 3 250 450 Sonifier Side View Dimensions 2 5 250 450 Sonifier Front View Dimensions 2 5 Converter Horn Assembly Stand Set ON OFF switch to PA HE 3 3 Methods For Connecting Removing Horn From Converter4 8 Connection or Removing and Horn 4 9 240 450 Sonifier Assembly Drawing 6 2 Soundproof Box 4 100 413 016 Rev 250 450 Sonifier User s Manual tab 2 1 tab 4 1 tab 4 2 tab 4 3 tab 5 1 tab 6 1 tab 6 2 tab D 1 100 413 016 Rev B List of Tables Front Panel Device and Function 2 2 Temperature Rise Variations 4 4 Horn Amplitude Table 4 10 Microtip Amplitudes 4 11 Symptom Probable Cause 5 1 Power Supply Parts 100 132 1622 6 3 Power Supply Parts 100 132 1623 6 4 Accessories Parts List 0 1 BRANSON 100 413 016 Rev B 250 450 Sonifier Chapter 1 Safety and Support User s Manual For Safety of Operating Personnel Chapter 1 Safety and Support 1 1 For Safety of Operating Personnel 1 1 1 2 About this Manual 1 1 1 3 Purpose of the Equipment 1 2 1 4 Components 1 2
7. Instruction Manual Irradiation on Various Biological Materials 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 ultrasonic 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 of reducing 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 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 s
8. Weight 4 Ibs 1 8 kg with horn Dimensions 7 L x 2 1 2 dia 180 x 60 mm Horn Diameter 1 8 1 3 2 25mm dia depending on process needs Power Supply Weight 15 Ibs 6 8 kg 2 3 3 Dimensions Figure 2 3 250 450 Sonifier Side View Dimensions fo 5 rH Hri 1 75 11 19 gt 16 83 gt 18 96 Figure 2 4 250 450 Sonifier Front View Dimensions 738 Duty w 8 ad BRANSON Power 100 413 016 Rev B Specifications BRANSON 2 6 100 413 016 Rev B 250 450 Sonifier Chapter 3 Delivery Handle User s Manual Unpacking and Handling Chapter 3 Delivery and Handle 31 Unpacking and Handling 3 1 3 2 Placement of Eguipment 3 1 3 3 Power Requirements 3 2 3 4 Setup Procedure 3 3 3 1 Unpacking and Handling Unpack the model 250 450 Sonifier unit as soon as it arrives Normal precautions 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
9. 10 ml solution completely disrupts 40 seconds The Sonifier Cell Disruptor has been used extensively in research on this organism Egg whites can be reduced to a homogeneous pipettable solution in 15 seconds on low power Ehrlich ascites disrupt in a few seconds Electron microscopy apertures are quickly cleaned A 4 100 413 016 Rev B 250 450 Sonifier Appendix The Effects of Ultrasonic Instruction Manual Irradiation on Various Biological Materials 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 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 fat can 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 Fluorocar
10. 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 3 2 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 on the bottom of the power supply is not blocked 100 413 016 Rev B 3 1 Power Requirements BRANSON 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 underneath the power supply to ensure that dust or dirt is not restrict ing the flow of air 3 3 Power Requirements The Sonifier unit requires a single phase three wire 117V or 200 245 V 50 60 Hz source The line cord is equipped with a NEMA 5 15P plug and requires a NEMA 5 15R receptacle To prevent the possibility of electrical shock always plug the Sonifier unit into a grounded power source 3 2 100 413 01
11. 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 100 413 016 Rev B A 1 The Effects of Ultrasonic Irradiation on Various Biological Materials BRANSON Aurefaciens monocellular elements from surface grown colonies obtained in 1 minute Complete disruption in 5 minutes 50 disruption in 2 minutes 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 of a 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 thermophillic spore form 98 disruption of 70 ml of 4096 suspension in 15 minutes Bacillus brevis 1 15 W V in 3 minutes Bacteroides Symbiosis 1 phosphofructokinase a sol
12. high meter deflection horn unloaded Power supply operates contin uously but does not operate in PULSED mode 1 Power supply module has failed 2 Horn or tip have failed 1 Pulse mode timer or circuitry failed A slight electric shock is felt when touching the Sonifier unit 1 Unit is improperly grounded 2 Line cord has failed 100 413 016 Rev B Chapter 5 General Information BRANSON 5 4 100 413 016 Rev 250 450 Sonifier Chapter 6 Main Electrical Assembly User s Manual Chapter 6 Main Electrical Assembly 250 450 Sonifier Assembly 6 2 6 2 250 Power Supply Parts List 6 3 6 2 1 Domestic 100 132 1622 6 3 6 3 450 Power Supply Parts List 6 4 6 3 1 Domestic 100 132 1623 6 4 6 4 250 450 Interconnect Schematic 6 5 100 413 016 Rev B 6 1 Chapter 6 Main Electrical Assembly BRANSON 6 1 250 450 Sonifier Assembly 100 244 005 100 244 006 Figure 6 1 240 450 Sonifier Assembly Drawing RF cable Line Filter Terminal Block Bracket Power Supply Capacitors Power Supply Module 6 2 100 413 01
13. is provided C 6 Soundproof Box Even though ultrasound is above the audible range of the 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 Soundproof Box 4 100 413 016 Rev 250 450 Sonifier Appendix D Accessories Parts List Instruction Manual Appendix D Accessories Parts List Accessories Parts List D 1 D 1 Accessories Parts List Table D 1 Accessories Parts List TTEM DESCRIPTION S 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 17 dia threaded body solid 101 147 044 100 413 016 Rev B D 1 Appendix Accessories Parts List BRANSON Table D 1 Accessories Parts List BRANSON TTEM DESCRIPTION EDP No 3 4 dia solid high gain 101 147 035 Cup Horns 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 Cup Horn High intensity with flow thru p
14. 1 5 Principle of Operation 1 3 1 1 For Safety of Operating Personnel 1 Make sure that the eguipment is properly grounded DO NOT operate if it is not 2 The unit is eguipped 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 any circumstances remove the power cord ground prong 3 DO NOT operate equipment with the cover removed High voltage is present within the equipment when connected to plant wiring 4 Do not allow the horn or micro tip to contact lab stands beakers etc or horn micro tip failure may result 5 Establish a standard of operation and periodically test equipment as described in Section 4 3 1 2 About this Manual This manual contains operation and service instruction for the Sonifier cell dis ruptor Model 250 450 It contains background information essential to the proper use and care of this equipment 100 413 016 Rev B 1 1 Chapter 1 Safety and Support Purpose of the Eguipment BRANSON The following definitions apply in this manual Inconvenience only if disregarded no damage or personal injury Eguipment damage may occur but not personal injury Personal may occur DO NOT DISREGARD 1 3 Purpose of the Eguipment The 250 450 Sonifiers can be used to disrupt cells bacteria spores or tissue and are ideal for initiating and accelerating chemical biochemical a
15. 250 450 Sonifier Analog Cell Disruptor User s Manual EDP 100 413 016 Rev B 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 B 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 Patents and Copyright Copyright 1998 by Branson Ultrasonics Corporation Danbury Con necticut U S A All rights reserved Branson Ultrasonics Corporation equipment is manufactured under one or more of the following U S Patents 4 973 876 5 435 863 5 855 706 Sonifier is a registered trademark of Branson Ultrasonics Corporation Danbury Connecticut U S A Other trademarks and service marks respectfully mentioned herein are held by their respective owners Loctite is a registered trademark of Loctite Corporation Newington CT Other trademarks and service marks mentioned herein are
16. 4 0 4 0 0 2 1 6 4 4 100 413 016 Rev B 250 450 Sonifier Chapter 4 Operation User s Manual Operating Considerations 4 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 lows 4 2 3 1 Foaming or Aerosoling Always place the horn deep enough below the surface of the liguid 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 vial such as a cut down Eppendorf tube is recommended The conical shape of this type of container effectively raises the level without increasing the volume thereby permitting the horn to be inserted more deeply below the liq 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 1 or 2 for a few seconds Then gradually advance the power output control to the level required 4 2 3 2 Tip Erosion Horn tips can erode Tip erosion is a side effect of the cavitation process that o
17. 6 Rev B 250 450 Sonifier User s Manual 6 2 250 Power Supply Parts List 6 2 1 Domestic 100 132 1622 Chapter 6 Main Electrical Assembly 250 Power Supply Parts List Table 6 1 Power Supply Parts 100 132 1622 Description Branson EDP No Designation Assembly Power Sup 100 244 126R ply Module Assembly R F Cable 8 100 241 345PR ft Assembly Line Cord 100 241 226PR 117V Filter Line 6A 200 163 033 200 126 008R Fl Fuse 3 AG 5 AMP 200 049 121R Meter Panel 100 068 018R SWI Switch ON OFF 200 099 296R Timer 200 104 009 SW 2 Switch Duty Cycle 200 099 170R 100 413 016 Rev B Chapter 6 Main Electrical Assembly 450 Power Supply Parts List 6 3 450 Power Supply Parts List 6 3 1 Domestic 100 132 1623 Table 6 2 Power Supply Parts 100 132 1623 BRANSON Description Branson EDP No Assembly Power Supply 100 244 125R Module Assembly R F Cable 8 ft 100 241 345PR Assy Line Cord 117V 100 241 226PR Filter Line 6A 200 163 033 200 126 008 FIF Fuse 3 AG 5 AMP 200 049 121R MI Meter Panel 100 068 018R SWI Switch ON OFF 200 099 296R SW 2 Switch Duty Cycle 200 099 170R 100 413 016 Rev B 250 450 Sonifier Chapter 6 Main Electrical Assembly User s Manual 250 450 Sonifier Interconnect Schematic 6 4 250 450 Sonifier Interconnect Schematic LINE 50 80Hz 230V 11
18. 6 Rev B 250 450 Sonifier Chapter 3 Delivery and Handle User s Manual Setup Procedure 3 4 Setup Procedure Figure 3 1 Converter Horn Assembly in Stand Set ON OFF switch to OFF a Plug the line cord intone appro priate electrical power outlet ensur ing that the power supply is grounded to prevent possibility of an electrical shock b Ensure that the horn is screwed into the converter and tightened adequately Remember to use washer in between Horn and Con verter The recommended torque is 220 inch pounds 24 85Nm c Mount converter horn assembly in laboratory stand or Branson sound enclosure as shown in Figure C 1 Secure clamp on converter d Ensure that the tip is screwed into the horn and tightened ade quately 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 4 100 413 016 Rev B 3 3 BRANSON 3 4 100 413 016 Rev B 250 450 Sonifier Chapter 4 Operation User s Manual General Operation Chapter 4 Operation 4 1 General Operation 4 1 4 1 1 Timed Operation 4 1 4 1 2 Continuous Operation 4 2 4 1 3 Pulsed Operation 4 2 4 2 Operating Considerations 4 3 4 2 1 Limiting Temperature Rise 4 3 4 2 2 Vessel Capacity and Speed of
19. 7V TERMINAL BLOCK BLU 2 2 BLU WHT BLU z Bree BLU JA ____ 5 T TLLWHT BLU i te BRN Iq BRN 1 1 1 yH HT BLU RFI 1 GRN s FILTER FILTER REAR PANEL 5432 1 21 CABLE c GRN YEL SND 54321 ASSEMBLY BLK 2424 o E RET sje BLK our OG RED GRN YEL WHT RED y eis cy sep om iiss atti fo of WHT BLU z Sl ml POWER SUPPLY m MODULE SEE NOTE 1 Nar USED 5 GRY MAN TUNE 3 MAN TUNE 2 MAN TUNE 1 RUN OVERLOAD INPUT OVERLOAD INDICATOR WHT YEL E NOT USED POWER SUPPLY CONTROL BOARD ODD NOO BU QONDU AU Q LO Q0 73 UT ADAPTER CONTROLLER PROGRAMMER MODEL B 1 51 51 Gi COMMON AMPLITUDE IN 10 REF AMPLITUDE OUT POWER FREQ OUT MEMORY FREQ OFFSET RUN NOTES UDNTU AWN VONGOUBUN 5 suni 1 PINS SHOWN ARE NOT USED
20. Temperature Rise 4 4 4 2 3 Minimizing Undesirable Factors 4 5 4 2 4 Sterilizing and Preventing Cross Contamination 4 6 4 2 5 Disrupting Tissues and Solids 4 6 4 2 6 Using Glass Powders with Solution 4 7 4 3 Testing the Equipment 4 7 4 4 Connecting and Removing Tips and Converters 4 8 4 4 1 Connecting Horn to Converter 4 8 4 4 2 Connecting to Horn 4 8 4 5 Horn and Micro Tip Amplitudes 4 10 4 1 General Operation In general starting conditions are as follows 4 1 1 Timed Operation Turn timer control clockwise to the desired processing time Set Duty Cycle Con trol to Constant and Output Control to 1 100 413 016 Rev B 4 1 General Operation BRANSON 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 4 1 2 Continuous Operation Turn Tlmer Control fully clockwise to the Hold position Set Duty Cycle Control to Constant and Power Switch to On Adjust Output Control to the desired required intensity 4 1 3 Pulsed Operation Use of the Duty Cycle control enables the ultrasonic energy to be pulsed
21. bons extended treatment time will break down particle size to well under 1 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 100 413 016 Rev B A 5 The Effects of Ultrasonic Irradiation on Various Biological Materials BRANSON 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 Herpes virus may be quickly released without injury Histoplasma capsulatum ultrasonic treatment for 7 minutes completely rup tured cells prepared by f
22. ccurs when liquids are submitted to ultrasonic energy The rate of erosion depends on the intensity of power applied corrosiveness of the liquid being treated and the amount of use Erosion accumulates increases and accelerates unless corrected in its early stages Recognize erosion early by periodically inspecting the tip Starting from a polished or shiny state the tip changes to light grey then dark grey then concen tric rings appear Finally the tip becomes very rough and pitted resulting in power output loss The tip can also discolor or darken the solution by introducing metal particles into it When the tip begins to turn to a medium grey color polish the grey area with a very fine crocus cloth To do this the tip or horn does not have to be removed from the converter place the crocus cloth on a flat surface hold the lower portion of the horn and rub it on the cloth for a few seconds to restore the original polish to the tip If the horn is also pitted use very fine emery cloth to remove the pits first 100 413 016 Rev B 4 5 Operating Considerations BRANSON 4 2 3 3 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 Place the horn in a container of alcohol and turn the power on for a few seconds to clean the interior th
23. d 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 Cell The Rosette Cell made of borosilicate glass has a conical shape with three side arms through which the solution is 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 B C 3 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
24. 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 Iysine 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 packed column 100 413 016 Rev B A 3 The Effects of Ultrasonic Irradiation on Various Bi
25. ed power supply tor light fails to illuminate when power supply is ener gized HF Line has failed Switch SI has failed Fuse FI has failed Line filter has failed to the horn No indication on meter Fan does not operate when 1 Fan motor has failed power supply 1s energized 2 Line cord failed Indicator light illuminates 3 Switch 51 has failed 4 Fuse has failed 5 Line filter has failed Fuse F1 fails when ON OFF 1 Underrated fuse switch is energized 2 Incorrect voltage 3 Fan motor has failed 4 Power Supply module has failed Ultrasonic power is delivered 1 Meter has failed 100 413 016 Rev B 250 450 Sonifier User s Manual Table 5 1 Chapter 5 General Information Symptom Probable Cause Symptom Probable Cause No or inconsistent ultrasonic power Some indication on meter 1 Foreign material between horn surface 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 no be immersed ON UA Little or no meter indication when ON OFF switch is ener gized Fan operates 1 Timer S3 has not been set has completed cycle or has failed 2 Thermo switch SW1 is acti vated 3 Pulsar board has failed Full scale or
26. erial 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 permanent 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 B 250 450 Sonifier How to obtain Further Assistance Instruction Manual Appendix How to obtain Further Assistance Customer Service Technical Support and Applications Assistance B 1 Equipment Failure Support 1 Returning Equipment 1 B 1 Customer Service Technical Support and Applications Assistance Customer Service 800 732 9262 Technical Support 800 732 9262 Applications Assistance 203 796 0532 B 2 Equipment Failure Support 203 796 0551 203 796 0355 203 796 0529 B 3 Returning Equipment Contact Custo
27. held by their respective owners 100 413 016 Rev B BRANSON Regulatory Compliance Sonifiers 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 EN61000 4 3 61000 4 6 and ENV50204 They take advantage of Branson latest System Protection and Autotune technology System protection constantly monitors various operating parameters of the power supply It if 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 condi tion has been sensed turn the power switch off and then turn it back n On Digital Sonifiers the test function will reset the power module Autotune optimizes the operating frequency of the power supply so that it is con sistent with the acoustic tooling To facilitate the auto tune process the power sup ply 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 fre quency The output meter on new CE 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 On CE Digital Sonifiers amplitude is now set digitally from the front panel similar to all other
28. itch to ON b Record meter reading with horn in air Meter Reading Horn Size c Fill 500 ml pyrex beaker to 500 ml level with room temperature tap water d 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 Section 5 100 413 016 Rev B 4 7 2 Tips Horns and BRANSON Converters 4 4 Connecting and Removing Tips Horns and Converters 4 4 1 Connecting Horn to Converter The 250 450 Sonifier horns have been torgued onto the converter at the factory Should you need 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
29. mer Service 800 732 9262 100 413 016 Rev B B 1 A z How to obtain Further BRANSON B 2 100 413 016 Rev B 250 450 Sonifier Appendix C Accessories Instruction Manual Appendix Accessories Micro Tip C 1 Cup Horn C 2 Flow thru Horn C 3 Rosette Cell O C 3 Continuous Flow Attachment C 4 Soundproof 4 Special disruptor horns various horn tips and a variety of vessels for batch 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 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 is 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 B C 1 Accessories BRANSON The stepped micro tip 1 a tw
30. minutes Mycobacteria a 20 ml growing media completely disrupts in 14 minutes Clumps break quickly An immunological compound prepared Mycoplasma antibody asuspension 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 3096 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 100 413 016 Rev B A 7 The Effects of Ultrasonic Irradiation on Various Biological Materials BRANSON Nocardia ostenodes breaks clumps and disintegrates in less than 10 minutes Nucleoprotein extracted from tissue May be degraded selectively 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 excelle
31. nd 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 B 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 1 4 Main Components The main components of the 250 450 sonifiers are the power supply the converter and the horn shown in figure 1 below 1 2 100 413 016 Rev B 250 450 Sonifier Chapter 1 Safety and Support User s Manual Principle of Operation Figure 1 1 Model 250 450 Sonifier Main Components 1 5 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 mechan ical vibrations The heart of the converter is a lead zirconate titanate electrostric tive element which when subjected to an alternating voltage expand and contracts The converter vibrates in a longitudinal direction and transmits the motion to the horn tip immersed in the solution The implosion of microscopic cavitation in the solution results causing the mol ecules in the medium to become intensely agitated The So
32. nifier 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 9 per second enabling a solution to be processed at full ultrasonic intensity while limiting temperature build up especially valuable when processing heat volatile solutions and temperature sen sitive material In Continuous Mode the Sonifier can be set up for continuous duty 100 413 016 Rev B 1 3 Principle of Operation BRANSON 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 ampli tude 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 ampli tude or with larger radiating surface is used or
33. nt 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 Pesticides ultrasonic treatment resulted in a 16 fold improvement in the potency of the 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 Pneumococci 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 A 8 100 413 016 Re
34. o piece horn consisting of a coupling section and lower tip Because the coupling section 18 attached directly to the converter the standard disruptor horn must be removed prior to using the stepped micro tip Recommended for use on extremely small volumes the 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 tissue culture cells HeLa cells and the complete range of cells which have low to medium resistance to breakage To prevent foaming or aerosoling while processing small quantities 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 7 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 Cup Horn at the top With chilled water in the Cup Horn the tubes are suspended in the Cup with the contents of the tubes just below the water level Ultrasonic energy is then conducted from the surface of the horn through the water and tube
35. ological Materials BRANSON Clostridium quickly disrupts all types Coagulase globulin ultrasonic treatment before precipitation yields signifi cantly more enzyme 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 5 minutes with 50 protein release and excellent enzyme activity Crypstostroma corticale maple bark spores a concentrated solution completely disrupts 1114 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 dispersion and homogenization coli 2 gm wet wt in
36. omogenizes in 2 minutes Lymphacytis complete disruption in 15 seconds Lymphocyte nuclei complete disruption in 6 minutes A 6 100 413 016 Rev B 250 450 Sonifier Appendix The Effects of Ultrasonic Instruction Manual Irradiation on Various Biological Materials Lymph gland direct injection lymphography with a modified radiopaque emulsion was obtained by ultrasonic treatment in a functional procedure produc ing lymphatic structure detail 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 2096 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
37. 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 if the process is not to be timed or a processing time of longer than 15 minutes is required Set Duty Cycle Control to the required pulse duration The lower number setting the shorter the pulse and the slower will be the temperature rise of the solution Switch power On and adjust Output Control to the required intensity 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 of the specimen using your estimated con trol variables When unsure initial test should be conduct with short time and low amplitude setting 4 2 100 413 016 Rev B 250 450 Sonifier Chapter 4 Operation User s Manual Operating Considerations 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 When using micro tips do n
38. onified 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 at short expo sure 100 413 016 Rev B A 11 The Effects of Ultrasonic Irradiation on Various Biological Materials 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 mat
39. operating parameters 100 413 016 Rev B 250 450 Sonifier User s Manual Warranty When used 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 Danbury Connecticut or an authorized repair station as advised by BRANSON provided the defective unit is returned properly packed with all transportation charges pre paid 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 product undue stresses that may damage the equipment Equipment failures resulting from using unqual ified horns and tips are not covered by the Branson warranty Microtips are designed to give maxim
40. ormalin fixation Good enzyme activity was obtained Human serum proteins ultrasonic treatment causes 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 Tysozyme activity The Iysozyme concentration of the leukocytes ug 10 6 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 h
41. orts 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 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 out breaking the airtight seal and permit processing with an inert gas Stainless steel 3 10ml 101 021 001 6 15ml 101 021 002 D 2 100 413 016 Rev B 250 450 Sonifier Instruction Manual Appendix D Accessories Parts List Table D 1 Accessories Parts List TTEM DESCRIPTION pon d 20 50ml 101 021 003 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 A
42. ot advance the output control beyond 7 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 the glass container can cause fracture of the tip or container 4 2 Operating Considerations The following sections discuss operating techniques under varying conditions 4 2 1 Limiting Temperature 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 Based on heat transfer characteristics the following vessel materials are recom mended listed in decreasing order of heat conductivity Best Aluminum Stainless Steel Thin wall Glass Thick wall Glass Poorest Plastic 100 413 016 Rev B 4 3 Operating Considerations BRANSON 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 i
43. pered 59 5 151 0 247 0 0023 0059 0108 101 063 212 Double Step 64 0 173 8 274 0 0025 0068 0108 All measurements in Microns 100 413 016 Rev B 4 11 5 aa BRANSON 4 12 100 413 016 Rev B 250 450 Sonifier Chapter 5 General Information User s Manual Chapter 5 General Information 5 1 General Information 9 1 5 1 1 System Trouble Analysis Chart 5 1 51 General Information If maintenance 15 reguired contact Branson Product Support Department Bran son Ultrasonics Corp Danbury Connecticut 1 203 796 0335 or 1 203 796 0551 If you have a problem operating your unit 1 Refer to the System Trouble Analysis Chart in this section to locate the symptom that most clearly describes your problem 2 Use the schematics and parts lists in Section 5 to identify parts or assemblies to be checked or replaced 3 Refer to the last page of the manual for equipment return informa tion 5 1 1 System Trouble Analysis Chart Table 5 1 Symptom Probable Cause Symptom Probable Cause Main power fuse fails or cir 1 Line cord is shorted cuit reader trips when system 2 Switch S1 has failed is plugged into electrical out 3 Line filter has failed let 100 413 016 Rev B 5 1 Chapter 5 General Information BRANSON Table 5 1 Symptom Probable Cause Symptom Probable Cause Fan does not operate Indica Unplugg
44. reads Remove the horn from the alcohol Operate the horn in air for a few seconds to dry the 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 1 4 20 tighten at 90 inch Ibs 10 16 Newton meters 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 4 2 3 4 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 4 2 4 Sterilizing and Preventing Cross Contamination Sterilize horns and tips by removing them from the converter and autoclaving them 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 4 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 tis
45. ries 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 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 B 250 450 Sonifier The Effects of Ultrasonic
46. s 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 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 B A 9 The Effects of Ultrasonic Irradiation on Various Biological Materials 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 laborato
47. s too great with this method consider using the following e ce salt 6 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 4 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 4 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 4 1 Temperature Rise Variations Sample Size 25 ml 100m 1 Power Level 3 7 10 3 7 10 Seconds Temperature after sonic time 30 307 359 429 26 5 27 5 29 3 60 342 459 55 6 127 59 30 3 33 5 120 429 61 78 30 35 41 3 180 48 5 74 90 32 39 3 48 240 54 59 82 59 95 34 44 54 5 300 60 88 959 36 48 60 Average Diff 0 7 0
48. sue 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 4 6 100 413 016 Rev B 250 450 Sonifier Chapter 4 Operation User s Manual Testing the Eguipment 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 4 2 6 Using Glass Powders with Solution To disrupt difficult cells 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 4 3 Testing the Equipment To establish a base line for future testing to verify if the equipment is operating properly proceed as follows Mount 1 2 disruptor horn with flat tip if tapped to converter a Set Output Control to 5 Timer to HOLD and ON OFF sw
49. t exchange Model 25 8 25 ml 201 123 001 Model 50 25 180ml 201 123 002 100 413 016 Rev B 250 450 Sonifier Instruction Manual Table D 1 Appendix D Accessories Parts List Accessories Parts List TTEM DESCRIPTION BRANSON EDP No Model 250 35 300ml 201 123 003 Tissue Disrup tor Designed for disintegration of difficult tissues Stainless steel construction Cell bot tom holds up to 6g of tissue A water jacket 1s provided for cooling 101 021 007 Soundproof Enclosure Reduces mechanical noise generated during liguid pro cessing to a normal level Especially useful when using a cell disruptor for extended periods 25 to 28 db cavita noise reduction 101 063 275 Glass Beads For cell homogenization Waterproof 1 pound packages 1 2 mm diameter 25 micron diameter 201 002 004 201 002 003 35 micron diameter 201 002 005 1 2 Wave Horn 100 413 016 Rev B Extends 1 2 dia disruptor horn an additional 1 2 wave length apprx 5 at a 1 2 diameter 101 147 049 Appendix D Accessories Parts List BRANSON D 6 100 413 016 Rev B
50. ton meters f 3 8 24 thread tighten at 180 inch Ibs 20 33 Newton meters Figure 4 2 Connection or Removing Tip and Horn 100 413 016 Rev B 4 9 Hon d io np AUGE BRANSON 4 5 Horn and Micro Tip Amplitudes Table 4 2 Amplitude Table Output Control Setting 1 5 10 101 147 037 1 2 Dia Tapped Stepped Disruptor 21 0 76 0 145 0 0008 003 0057 Part No Description 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 0027 0051 101 147 044 1 0 Dia Solid Stepped 6 3 121 5 40 5 0002 0008 0016 All measurements in Microns 4 10 100 413 016 Rev B 250 450 Sonifier User s Manual Chapter 4 Operation Horn and Micro Tip Amplitudes Table 4 3 Microtip Amplitudes Part No Description Output Control Setting 1 5 10 101 148 062 1 8 Dia Tapered 116 0 306 0 494 0 0046 012 0194 101 148 069 3 16 Dia Tapered 59 5 183 0 302 0 0023 0072 0119 101 148 070 1 4 Dia Ta
51. ttaches 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 101 148 070 100 413 016 Rev B D 3 Accessories Parts List Table D 1 BRANSON Accessories Parts List TTEM BRANSON DESCRIPTION EDP No Double Step Micro Tip Assembly A two piece horn consisting of a coupling sec 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 Coupler Section Only 101 147 050 Micro Tip Section Only 101 148 063 Stud Replacement adaptor stud 100 098 249 Horn to Converter Rosette Cool ing Cell 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 hea
52. uble 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 require up to a week Bone specimens treated in this way yielded large numbers of intact cells with little distortion Malignant criteria were easily recognized Tumor types studied were osteosarcoma chondrosar coma liposarcoma chordoma metastic bronchogenic squamous 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 require extensive treatment time A 2 100 413 016 Rev B 250 450 Sonifier Appendix The Effects of Ultrasonic Instruction Manual Irradiation on Various Biological Materials 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 Brevi bacterium 25 ml disrupts in 20 seconds Brevi bacterium acetylicum approximately 3 minutes to
53. um mechanical energy output Since they operate close to the stress limits of titanium 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 B BRANSON 100 413 016 Rev B 250 450 Sonifier User s Manual Table of Contents 1 Safety and Support 1 1 1 1 For Safety of Operating Personnel 1 1 1 2 About this Manual 1 1 1 3 Purpose of the Equipment 1 2 1 4 Components 1 2 1 5 Principle of Operation 1 3 2 Introduction 2 1 2 1 250 450 Sonifier Power Supply Front Panel 2 1 2 2 Model 250 450 Sonifier Power Supply Rear Panel 2 3 2 3 Specifications 2 4 2 3 1 Electrical Specifications 2 4 2 3 2 Mechanical Specifications 2 5 2 3 3 Dimensions 2 5 3 Delivery and Handle 3 1 31 Unpacking and Handling 3 1 3 2 Placement of Equipment 3 1 3 3 Power Requirements 3 2 3 4 Setup Procedure
54. v B 250 450 Sonifier A The Effects of Ultrasonic Instruction Manual Irradiation on Various Biological Materials Pseudomonas fluorescens 2 gm wet wt in 10 ml completely disrupts in 1 minute Pulmonary cytodiagnosis the mucous in sputum 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 element
55. 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 two types of Cup Horns a high intensity unit that will accommodate a single test tube and a larger unit for multiple test tubes The high intensity Cup Horn has a concave bottom that focuses the ultrasonic energy on the bottom of the tube The larger unit with a diameter of 2 or 3 inches allows the immersion of mul tiple 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 Horns C 2 100 413 016 Rev B 250 450 Sonifier Instruction Manual are designed to allow chilled water to circulate through the Cup Horn to prevent heating of the solutin as result of the ultrasonic activity The Cup Horn is avail able 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 is ultrasonically activate
56. when any horn is driven at higher amplitude by increasing the output control setting 1 4 100 413 016 Rev B 250 450 Sonifier Chapter 2 Introduction User s Manual 250 450 Sonifier Power Supply Front Panel Chapter 2 Introduction 2 1 250 450 Sonifier Power Supply Front Panel 2 1 2 2 Model 250 450 Sonifier Power Supply Rear Panel 2 3 2 3 Specifications 2 4 2 3 1 Electrical Specifications 2 4 2 3 2 Mechanical Specifications 2 5 2 3 3 Dimensions 2 5 2 1 250 450 Sonifier Power Supply Front Panel Figure 2 1 Supply Front Panel 4 BG 9 BRANSON id 2 SONIFIER al L lm 3l QD 5 31 4 Both illustrations represent Sonifier Power Supply Front Panel with the exception that the image shown as is intended for use in Japan 100 413 016 Rev B 2 1 Chapter 2 Introduction 250 450 Sonifier Power Supply Front Panel BRANSON Table 2 1 Front Panel Device and Function 1 2 Duty Cycle dial 3 ON OFF Switch 4 Loading Meter 5 Output control Device and Function Times the application of ultrasonic energy 0 to 15 minutes or to HOLD Use HOLD in either the PULSED or CONTINUOUS Modes if you require an undetermined e
57. xposure time and ultrasonic energy will be on indefinitely Set the timer any where from 0 to 15 minutes to either PULSED or CONTINUOUS Modes and ultrasonic energy will switch off at the end of the predetermined period 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 sec ond if you choose a 90 setting ultrasonics will be on for 90 of every second Applies main power to the unit Lights when power is on Indicates level of ultrasonic power delivered to your sample Controls the amplitude i e peak to peak motion of the ultrasonic vibrations Clockwise rotation increases amplitude For further information refer to Figures 6 and 7 Power Outputs Charts When using micro tips maximum output control setting is 7 100 413 016 Rev B 250 450 Sonifier Chapter 2 Introduction User s Manual Model 250 450 Sonifier Power Supply Rear Panel 2 2 Model 250 450 Sonifier Power Supply Rear Panel Figure 2 2 Power Supply Rear Panel BRANSON Table 22 Rear Panel Device and Function Item Device and Function 1 Power Connects the power supply to an electrical outlet Cord 2 Fuse 4 Overload Protection AMP 3 6 Pin Accessory connector
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