Cyto Pulse Sciences, Inc.
Contents
1. Cyto Pulse Sciences Inc Model PA 301 Pulse Booster User Manual Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 1 410 787 1890 1 410 787 1891 FAX e mail customer cytopulse com Cyto Pulse Sciences Inc makes no warranty with respect to the product except for the warranty set forth in this Users Manual on page 5 1 The LIMITED WARRANTY SET FORTH ON PAGE 5 1 IS EXCLUSIVE AND NO OTHER WARRANTY WHETHER WRITTEN OR ORAL IS EXPRESSED OR IMPLIED Cyto Pulse Sciences Inc SPECIFICALLY DISCLAIMS IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE License Agreement PA 301 Pulse Booster Cyto Pulse Sciences Inc Licensor conveys to the licensee for fee paid a nonexclusive nontransferable license to use the PulseAgile hardware and software equipment for research purposes into perpetuity Cyto Pulse Sciences Inc has patent allowance and patents pending covering the PulseAgile and other processes If the licensee wishes to use the hardware and software for production or commercial purposes an additional license shall be required The equipment is not approved by the FDA for use with in vitro or in vivo diagnostics or therapy The information in this manual is subject to change without notice Copyright 2000 2005 Cyto Pulse Sciences Inc PA301UMANrev 1 11 04 Price 100 00 PA 301 User Manual Rev 1 12 04 Cyto Pulse Sciences Inc Model PA 301 Pulse Booster U2. e Connect the Option Interconnect cable between the DB25 jacks on the back of the PA 301 and PA 4000 IDO NOT ATTACH TO A COMPUTER PARALLEL PORT OR A PRINTER e Connect the high voltage cable between the MHV jack on the back of the PA 4000 and the MHV jack labeled Pulse In on the back of the PA 301 This cord delivers the pulses generated by the PA 4000 to the PA 301 IDO NOT USE A CABLE WITH BNC PLUGS e Connect the HV cable with the MHV plug from 4 the CE 20 to the MHV jack labeled Pulse Out Figure 2 2 Back Panel Connections SHINE Dace OMNE FAGOT PArAUUU PASH 1DO NOT CONNECT A PA 96W TO THIS JACK e Connect the Cuvette Interlock cable from the CE 20 to the RCA type phono jack on the back panel of the PA 4000 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 2 1 PA 301 User Manual Ch2 Rev 1 12 04 2 3 PA 301 Front Panel Display The PA 301 front panel display consists of two light emitting diodes LED Both of the LEDs will light when the PA 301 is properly connected to the PA 4000 and line mains power is turned on The nterface LED indicates that the DB25 interface cable is installed and communication between the PA 4000 and the PA 301 has been established The Power LED indicates that the PA 301 line mains cord is plugged into the back of the PA 4000 and that 2 PA i ihe PA 301 is receiving power Figure 2 3 PA 301 Front Panel Display 2 4 Power Up With all cables connected
3. 80 C Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 PA 301 User Manual Ch3 Rev 1 12 04 4 3 3 E Coli Transformation Example The following procedure was performed to transform E coli with a plasmid containing a Kanamycin resistance gene 1 9 10 Prepared culture plates in advance Mix LB agar and water Microwave to boiling Microwave on low 1 minute Cool to 70 C Add Kanamycin if desired 10 l mil of 100 ug ml Kanamycin Pour 13 mL into 100 mm petri dishes and cool to room temperature Thawed 1 vial of Invitrogen Electromax DH5a E cells Added 2 uL plasmid 2 5 pg mL to 100 uL cells Note as little as 20 uL cells can be used Added cells to a 1 mm gap cuvette Applied three pulses 2000 volts amplitude 100 us pulse width 0 2 second interval using a Cyto Pulse Sciences PA 4000 PA 301 Pulse Booster system Added 1 ml SOC medium mix well and moved to Falcon tube Prepared 10 fold dilutions Plated dilutions by adding 5 ul cell suspension to culture plate with Kanamycin and to an identical plate without Kanamycin Incubated 18 hours 37 C Counted colonies and analyzed selected colonies using an Invitrogen Clone Checker System Results 130 000 transformed colonies were obtained 65 colonies counted at 1 100 dilution assaying 1 20 of the cell suspension References Nickoloff editor Methods in Molecular Biology Vol 47 Miller EM Nickoloff JA Chapte
4. 1000 volts not 3000 volts This is the case because the PA 4000 is going to deliver a 1000 volt pulse to the PA 301 which in turn will boost the pulse voltage by a factor of three subsequently delivering a 3000 volt pulse to the cuvette Click Start The log report should look something like that shown in Figure 3 3 Protocol File PA 301 Test pro 12 7 2004 3 13 59 40 Mode Electroporation w 3 1 Transformer Monitor Voltage x 3 gt GRP NUM WIDTH INTVL gt 1 1 0 100 0 50 gt GRP NUM WIDTH INTVL gt 2 1 0 200 0 50 gt GRP NUM WIDTH INTVL gt 3 1 0 400 0 50 gt GRP NUM WIDTH INTVL gt 4 1 0 600 0 50 gt Estimated load gt 100 ohms gt Estimated conductance lt 0 001 siemens Figure 3 3 PA 301Test pro Protocol Log Report mportant to note that when programming a protocol the user will enter the pulse amplitude of the waveform delivered to the sample That is the pulse out of the PA 301 and to the test sample The PulseAgile software however is monitoring the voltage set inside the PA 4000 which will be one third of the output pulse This concept is illustrated in Figure 3 4 User programs PA 4000 delivers 3000 Volt pulse 1000 Volt pulse to PA 301 PA 301 delivers 3000 Volt pulse to cuvette PA 4000 Figure 3 4 Relationship among User programmed volts PA 4000 volts and PA 301 volts Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 PA 301 User Manual Ch3 R
5. Pulse Width 0 001 to 0 600 ms in 0 001 ms steps 0 100 ms maximum at 3000 V Minimum Load 200 ohms This manual has been designed to help you realize the maximum benefit from using the PA 4000 PA 301 PulseAgile Pulse Booster system It contains information on how to operate the electroporator safety tips and applications Note The PA 4000 with the PA 301 contains a high voltage power supply and was designed with safety features to protect the user and the equipment If used properly the PA 4000 with the PA 301 is a safe and reliable instrument Chapter 2 explains some important concepts related to operator safety in addition to concepts needed for accurate use of the instrument Chapter 2 must be read before setting up this instrument Our goal is the safe and productive use of the PA 4000 with the PA 301 Back Panel Symbols OA A 3 Protective Caution Caution Chassis Terminal Refer to Risk of Ground Conductor documentation electric shock The PA 4000 with the optional PA 301 is rated for operation with line mains voltage of 100 240 VAC maximum current of 2 amps at 50 60 Hz The AC mains power supply cord is the disconnect device for this product The power supply cord shall be a Type SJT rated 300 Volts AC 18 AWG 105 C 3 conductor including ground This unit is rated for environmental conditions of 5 40 C 80 relative humidity to 31 C decreasing linearly to 50 relative humidity at 40 C altitude to 2000 meters
6. including the serial communication cable from the host computer turn on line mains power and start the PulseAgile software The set up of the system is now complete The Getting Started chapter will give instruction on the proper operation of the Pulse Booster System 9 29 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 PA 301 User Manual Ch3 Rev 1 12 04 3 Getting Started with the PA 301 3 1 Introduction The PA 301 Pulse Booster System expands the high voltage pulse amplitude range available to researchers It is capable of producing rectangular wave pulses up to 3000 A summary of the specifications is provided below This Chapter will provide details regarding the proper operation of the PA 301 using the PA 4000 PulseAgile interface software 3 2 Application Software The PA 301 application software is a part of the PA 4000 PulseAgile interface software It is accessible only when a PA 301 is connected using the DB25 interface cable as described in Chapter 3 After installing the PA 301 close the Cuvette Holder handle turn on the PA 4000 and start the PulseAgile software The screen should appear as shown in Figure 3 1 In the Options Connected area look for a check mark in the box labeled 3 1 Transformer If there is no check then the DB25 interface cable is not connected properly Power down the system and check the cable If the problem persists then call Cyto Pulse Sciences Customer Se
7. 1 User Manual Rev 1 12 04 Caution Notice The PA 4000 to be connected to this instrument contains a high voltage power supply that can be adjusted to beyond 1 000 volts Additionally the PA 301 can output a pulse of 3 000 volts Such voltages can be lethal The user must read this manual carefully before the instrument is placed into operation Removing the cover may void the warranty Do not connect or disconnect the high voltage cable with the high voltage enabled To connect or disconnect the cable turn line mains power off and unplug line cord Do not open the cuvette holder while the high voltage is on If a problem occurs during a run push the STOP RESET button on the front panel If there are any questions about the operation of this instrument call Cyto Pulse Customer service Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 787 1890 v PA 301 User Manual Rev 1 12 04 vi Blank Page Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 787 1890 PA 301 User Manual Ch1 Rev 1 12 04 1 Introduction The PA 301 Pulse Booster add on to the PA 4000 computer controlled electroporator provides the ability to establish higher electric fields required for smaller prokaryotic cells All pulse parameters are tunable to give the operator maximum flexibility in performing protocols Summary of PA 4000 PA 301 Pulse Booster System Pulse Specifications Pulse Amplitude 300 to 3000 V in 15 V steps
8. MD 21045 410 787 1890 PA 301 User Manual Rev 1 12 04 Table of Contents Introduction PA 4000 PA 301 Set up and Operation 2 1 Introduction 2 2 Connecting the PA 301 to the PA 4000 2 3 PA 301 Front panel display 2 4 Power Up Getting Started With the PA 301 3 1 Introduction 3 2 Application Software 3 2 1 3 2 2 Electroporation Protocol Screen Protocol programming 3 2 2 1 Programming Pulse Amplitude 3 2 2 2 Programming Pulse Width 3 2 3 Run a protocol PA 301Test pro Applications 4 1 Introduction 4 2 Electric Field Intensity 4 2 1 Rectangular vs Exponential Decay Waveforms 4 3 Preparation of Electrocompetent Cells 4 3 1 4 3 2 4 3 3 Customer E coli and Gluconobacter Lactococcus E Coli Transformation Example Service 4 1 Limited Warranty 4 2 Customer Service Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 787 1890 Page 1 1 2 1 2 1 2 3 3 1 3 1 4 1 5 1 PA 301 User Manual Rev 1 12 04 List of Figures The Pulse Booster Electroporation System Back Panel Connections PA 301 Front Panel Display PulseAgile Opening Screen showing PA 301 Pulse Booster attached PulseAgile Electroporation Protocol Programming Screen PA 301Test pro Protocol Log Report Relationship among User programmed volts PA 4000 volts and PA 301 volts Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 787 1890 Page 2 1 2 1 2 2 3 1 3 2 3 4 3 4 PA 30
9. R OTHER USES OF THIS PRODUCT CYTO PULSE S OBLIGATION TO REPAIR OR TO REPLACE TO THE EXTENT SET FORTH ABOVE CONSTITUTES THE EXCLUSIVE REMEDIES OF THE CUSTOMER FOR ANY BREACH OF WARRANTY This warranty shall not apply to products which after inspection by CYTO PULSE were found to be improperly used or to have been modified in any manner CYTO PULSE recommends that the user not open the product cabinet This limited warranty is valid for two years from the date of shipment 5 2 Customer Service If the user believes that there is a defect in the CYTO PULSE product the customer should contact CYTO PULSE Customer Service by e mail at customer cytopulse com or phone 410 787 1890 or contact the local CYTO PULSE representative A determination if the product is still in warranty will be made If the warranty period is still in effect the user will be given an authorization number RMA to return the product If after receipt and inspection the product is found to be defective it will be replaced or repaired and returned to the customer If the product is found to have been modified or misused the user will be given a quote for repair If the warranty period has expired and the user requests repair CYTO PULSE will inspect the product and provide a written quote for repair The user must provide a purchase order number before the product will be repaired If the unit is damaged in shipment the user must recover the insured value to replace or repair
10. There are no operator replaceable parts inside the system Cyto Pulse recommends that the user not remove the cabinet covers Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 715 0990 1 1 PA 301 User Manual Ch1 Rev 1 12 04 Blank Page 1 2 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 715 0990 PA 301 User Manual Ch2 Rev 1 12 04 2 PA 4000 PA301 Set Up 2 1 Introduction The PA 301 Pulse Booster is an optional attachment to the PA 4000 PulseAgile Electroporator It is a transformer that boosts the pulsed voltage output of the PA 4000 by a factor of three Subsequently the pulsed electric field produced in a cuvette is 3 times that produced by the PA 4000 alone For example electric field stenge of up to 7500 V cm are capable in a standard 4 mm cuvette The complete Pulse Booster system consists of PA 4000 Electroporator PA 301 Pulse Booster PA4 SW PulseAgile Software CE 20 Cuvette Holder CS LN Line Cable set CS OPT Interface Cable Set CUV M Cuvette Multi Pack PA4 UMAN PA 4000 User Manual PA301 UMAN PA 301User Manual Laptop shown is not included Figure 2 1 The PA 4000 PA 301 Pulse Booster Electroporation System 2 2 Connecting the PA 301 to the PA 4000 The PA 301 should be placed on top of the PA 4000 The Mains Line Power Switch must be off and the Mains Line Power Cord must be unplugged from the PA 4000 Referring to Figure 2 2 make the following connections
11. aximum allowable pulse width in the PA 301 Pulse Booster System varies according to the desired pulse amplitude This insures that all pulses delivered meet Cyto Pulse Sciences standard of pulse droop of lt 5 The table below gives the maximum pulse width allowed for ranges of pulse amplitude settings Pulse Amplitude Maximum Pulse Width volts ms 300 585 0 600 600 795 0 500 810 1095 0 300 1110 1590 0 200 1605 2190 0 150 2205 3000 0 100 If values outside the above ranges are entered the software will display an error message window and then set the pulse width to the maximum allowed value as demonstrated here Click on the Pulse Width parameter box Change the value to 1 000 ms Click Replace The following Message Box should display Pulse Agile Interface 2 0 x For an amplitude of 315 volts the maximum allowed pulse width is 0 6 ms Click OK and note how the Pulse Width is now set to 0 600 ms Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 3 3 PA 301 User Manual Ch3 Rev 1 12 04 3 2 3 Runa protocol PA 301Test pro Before running this protocol check that there is a cuvette installed in the cuvette holder and the handle is closed For this test an empty 4mm or greater cuvette is recommended It is i Open the protocol file PA 301 Test pro located in the protocol folder Click Turn HV On After the system charges note that the Power Supply Voltmeter reads
12. ev 1 12 04 4 Applications 4 1 Introduction The general principles for electroporation of prokaryotic organisms are the same as those described for eukaryotic organisms in the PA 4000 User Manual However the smaller size of the prokaryotic cells and the presence of a cell wall around many prokaryotic cells present challenges to efficient transfection through electroporation 4 2 Electric Field Intensity Electric field intensity is the most important pulse parameter determining electroporation efficacy Since the required electric field is inversely proportional to cell diameter smaller cells require higher electric fields There is a wide range of electric field intensity 3 30 KV cm reported in the literature for electroporation of small prokaryotic cells most are for E coli The majority of protocols use electric fields around 12 KV cm In contrast the electroporation of eukaryotic cells is generally performed at electric field intensities on the order of 1 kV cm to 2 5 kV cm 4 2 1 Rectangular vs Exponential Decay Waveforms Most published methods for electroporation of E coli describe exponential decay pulses The PA 4000 with PA 301 delivers a rectangular wave pulse There is no direct way to convert an exponential decay pulse protocol to a rectangular wave pulse protocol For best results a range of parameters should be tried initially Here are guidelines that can be followed to assist in making that conversion 1 Recta
13. from the carrier Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 5 1 PA 301 User Manual Ch5 Rev 1 12 04 Blank page 5 2 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990
14. g procedures can be used 4 3 1 1 OV Re GO NS 10 11 12 13 4 3 2 0 200 INE Oe OW o 4 2 E coli and Gluconobacter Grow 1 Liter culture to Asoo of 0 5 This usually requires growing a small culture overnight from a colony then inoculating two 500 ml cultures on a shaker at 37 C Add culture to 500 ml centrifuge tubes and cool to 4 C Centrifuge at 3000 X g for 10 minutes Discard supernatant and add 250 ml ice cold water or 1 mM HEPES pH 7 0 Centrifuge at 3000 X g for 10 minutes Discard supernatant and add 40 ml ice cold water or 1 mM HEPES pH 7 0 and transfer to 50 ml conical tube Centrifuge at 3000 X g for 10 minutes Discard supernatant and add 20 ml ice cold10 glycerol in water Centrifuge at 3000 X g for 10 minutes Aspirate supernatant and combine all pellets in 1 5 ml cold 10 glycerol in water Aliquot in small amounts 40 to 80 ul Freeze in liquid nitrogen or an ethanol dry ice bath Store at 80 C Lactococcus Grow L lactis in GM17 30 C to A600 of 0 5 Dilute cells and grow in SGM17 containing 0 2 to 4 glycine strain dependent to A600 of 0 5 Centrifuge 5000 X g 4 C Discard supernatant and add ice cold10 glycerol in 0 5 M sucrose Centrifuge 5000 X g 4 C Discard supernatant and add ice cold10 glycerol in 0 5 M sucrose Centrifuge 5000 X g 4 C Discard supernatant add 1 100 culture volume of ice cold 10 glycerol in 0 5M sucrose Use immediately or store at
15. ngular wave pulses are more precisely controlled than exponential decay pulses For instance the pulse width can be fine tuned to avoid overheating 2 Since rectangular wave pulses are shorter in duration than exponential decay pulses a higher pulsed electric field may be required For example a protocol designed around a 12 KV cm field delivered by a exponential decay pulser might require that a 20 KV cm field be used if redesigned for a rectangular wave pulser The Pulse Booster system can produce a field intensity of up to 30 KV cm in a 1mm cuvette 3 Multiple rectangular wave pulses may be better than single pulses 4 Since less heat is generated using rectangular wave pulses the cells should not be pre cooled Electroporation should be done at room temperature 5 More cells will be viable after using rectangular wave electroporation Rectangular and exponential decay waveform electroporation do have some common requirements however e Low conductivity medium should be used to reduce arching and heat e Electroporation competent E coli or other bacteria should be used Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 4 1 PA 301 User Manual Ch3 Rev 1 12 04 4 3 Preparation of Electrocompetent Cells Bacterial cells need to be prepared for electroporation by making them electrocompetent Electrocompetent cells can be purchased from companies such as Invitrogen Alternatively one of the followin
16. r 8 Echerichia coli Electrotransformation 1995 105 113 Mostofa HE Heller KJ Geis A Cloning of Escherichia coli lacz and lacY genes and their expression in Gluconobacter oxydans and Acetobacter liquifaciens Applied and Environmentqal Microbiology 2002 68 5 2619 2623 Nickoloff editor Methods in Molecular Biology Vol 47 Chapter 19 Transformation of Lactococcus by electroporation 1995 195 199 1 2 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 4 3 PA 301 User Manual Ch3 Rev 1 12 04 Blank Page 4 4 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 PA 301 User Manual Ch5 Rev 1 12 04 5 Customer Service 5 1 Limited Warranty CYTO PULSE products are warranted against defect in materials and workmanship If the customer provides notice of such a defect during warranty period CYTO PULSE at its option will either repair or replace the products which were found to be defective The limited warranty set forth above is exclusive and no other warranty whether written or oral is expressed or implied CYTO PULSE specifically disclaims implied warranties of merchantability and fitness for a particular purpose EXCEPT AS SET FORTH ABOVE CYTO PULSE MAKES NO WARRANTY WITH RESPECT TO THE PRODUCT AND IN NO EVENT REGARDLESS OF CAUSE SHALL CYTO PULSE BE LIABLE FOR INDIRECT SPECIAL OR CONSEQUENTIAL DAMAGES OR OTHER LOSSES OF ANY KIND ARISING FROM BREACH OF WARRANTY O
17. rvice PA 4000 Interface Version 2 01 E x File Tools Settings Help Options Connected ELECTROPORATION ryoroMmaera 7 3 1 Transformer System CormmLink k Electrode Holder OFF High Voltage Status Local Remote Monitors Power Supply Voltmeter volts Load Ohmmeter ohms Figure 3 1 PulseAgile Opening Screen showing PA 301 Pulse Booster attached Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 3 1 PA 301 User Manual Ch3 Rev 1 12 04 3 2 1 Electroporation Protocol Screen Click Electroporation and the screen should now appear as shown in Figure 3 2 The default pulse parameters are shown in the various boxes Details regarding programming protocols were given in Chapter 4 of the PA 4000 User Manual so this chapter will only cover the software functions specific to the use of the PA 301 3 2 2 Protocol programming There are several differences between programming a Pulse Booster protocol and a standard PA 4000 protocol Specifically the minimum and maximum allowable pulse amplitudes are different and the maximum allowable pulse width varies with pulse amplitude range Additionally there is no Low Range High Range select the PA 4000 reservoir capacitor is fixed to the High Range value by default 3 2 2 1 Programming Pulse Amplitude The minimum pulse amplitude for the Pulse Booster system is 300 volts As an exercise try this e Click on the Pulse Amplitude parameter bo
18. ser Manual Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 1 410 787 1890 1 410 787 1891 FAX e mail customer cytopulse com Cyto Pulse Sciences Inc makes no warranty with respect to the product except for the warranty set forth in this Users Manual on page 5 1 The LIMITED WARRANTY SET FORTH ON PAGE 5 1 IS EXCLUSIVE AND NO OTHER WARRANTY WHETHER WRITTEN OR ORAL IS EXPRESSED OR IMPLIED Cyto Pulse Sciences Inc SPECIFICALLY DISCLAIMS IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE License Agreement PA 301 Pulse Booster Cyto Pulse Sciences Inc Licensor conveys to the licensee for fee paid a nonexclusive nontransferable license to use the PulseAgile hardware and software equipment for research purposes into perpetuity Cyto Pulse Sciences Inc has patent allowance and patents pending covering the PulseAgile and other process If the licensee wishes to use the hardware and software for production or commercial purposes an additional license shall be required The equipment is not approved by the FDA for use with in vitro or in vivo diagnostics or therapy The information in this manual is subject to change without notice Copyright 2000 2005 Cyto Pulse Sciences Inc PA301UMANrev 1 11 04 Price 100 00 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 410 787 1890 i PA 301 User Manual Rev 1 12 04 Blank Page ii Cyto Pulse Sciences Inc P O Box 609 Columbia
19. x e Change the value to 100 volts e Click Replace e Note how the Pulse Amplitude defaults back to 300 volts PA 4000 Interface Version 2 01 olx Fie Tools Settings Help eLectroporation T ME 00 80 Bsa o Sas Canteen I AC Ger 7 3 1 Transformer GROUP ID GROUP LIST System PULSES CommLink 5 Jor Electrode Holder Amplitude ry 200 a High Voltage kA fi ead Status Local Remote l smote Monitors Power Supply Show Log Duration Peaksa A RS E EE vois History o 100 ms ae aed Ohmmeter ohms Last Protocol Log z Figure 3 2 PulseAgile Electroporation Protocol Programming Screen 3 2 Cyto Pulse Sciences Inc P O Box 609 Columbia MD 21045 USA 410 715 0990 PA 301 User Manual Ch3 Rev 1 12 04 The maximum pulse amplitude for the Pulse Booster system is 3000 volts So now try this Click on the Pulse Amplitude parameter box Change the value to 3100 volts Click Replace Note how the Pulse Amplitude defaults to 3000 volts The minimum voltage step achievable is 15 volts for a standard PA 4000 it is 2 volts in Low Range and 5 volts in High Range Here is an exercise to demonstrate this Click on the Pulse Amplitude parameter box Change the value to 310 volts Click Replace Note how the Pulse Amplitude defaults to 300 volts Change the value to 320 volts Click Replace Note how the Pulse Amplitude defaults to 315 volts 3 2 2 2 Programming Pulse Width The m
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