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Gemini Series Electroporator User’s Manual

toll free 800.272.2775 • 508.893.8999 • www.btxonline.com

21

Double tap New Protocol.

Use keyboard to name protocol.

Tap the Green Check icon to save.

Performing Experiments

Creating New Protocols

On the home screen,  tap User Protocols icon.

Creating an Exponential Decay Wave Protocol

Tap the Exponential icon.

Tap the Green Check icon to save.

Double tap your newly saved protocol.

Tap the Voltage icon.

Use the numeric keypad to set voltage.

Tap the Green Check icon to save.

PROTO SET TYPE

EDIT SCREEN

 

 SET VOLUME

Summary of Contents for Gemini SC

Page 1: ...Gemini Series Twin Waveform Electroporation Systems User s Manual www btxonline com 5507 002 REV 1 0 ...

Page 2: ... of the product Do Not Operate with Suspected Failures If damage is suspected on or to the product do not operate the product Contact qualified service personnel to perform inspection Orient the Equipment Properly Do not orient the equipment so that it is difficult to operate the disconnection device Place Product in Proper Environment Review the operating manual for guidelines for proper operatin...

Page 3: ...hnical support team constantly tracks published literature for any reference to electroporation and electrofusion We extract the pertinent experimental conditions and yields from these papers to help us in our efforts to help you In addition to tracking publications we are available to you for support at any time for advice in experimental design product recommendations troubleshooting and any oth...

Page 4: ... Backlight Preferences 17 Displaying Device Information 17 Preset Protocols 18 20 Using Preset Protocols 18 Customizing a Preset Protocol 19 20 Performing Experiments 20 26 Creating New Protocols 21 Creating an Exponential Decay Wave Protocol 21 23 Creating a Square Wave Protocol 24 26 Using Specialty Protocols 27 29 Using an HT Plate Handler 27 28 Using Specialty Electrodes 28 29 Managing Protoco...

Page 5: ...ate cells in suspension in cuvettes This system cannot accommodate multiple pulsing with the exponential decay waveform remote operation specialty electrodes or 96 well options For this reason some of the specifications of the Gemini SC are not as extensive as what is available in the Gemini X2 The BTX Gemini X2 is a highly advanced twin waveform generator incorporating both square and exponential...

Page 6: ...on number and inter pulse interval Research shows that certain experimental conditions and parameters of electrical pulses may be capable of causing many more molecules to move per unit time than simple diffusion There is also good evidence Sukharev et al 1992 that DNA movement is in the opposite direction An additional important consideration is when the voltage pulse is applied to the cells and ...

Page 7: ...ore formation to occur in the cell membrane Three factors should be considered for optimizing field strength 1 Electrode Gap Size 2 Cell Diameter 3 Temperature Cell Type Field Strength Ranges Bacteria Yeast 3 24 kV cm Mammalian 0 25 3 kV cm Plant 3 12 kV cm Electrode Gap Size The distance between electrodes or gap size is important when optimizing your electroporation experiment Field strength is ...

Page 8: ...ia which may contain serum Other recommended buffers are hypoosmolar buffers in which cells absorbs water shortly before pulse This swelling of the cells results in lowering the optimal permeation voltage while ensuring the membrane is more easily permeable for many cells but can be damaging to others Prokaryotic cells such as bacteria require the use of high resistance buffers 3000 ohms for this ...

Page 9: ... Restifo Steven A Rosenberg and RichardA Morgan High efficiency transfection ofprimary human and mouse Tlymphocytes using RNA electropora tion Molecular Therapy 2006 13 151 159 William J Buchser Jose R Pardinas Yan Shi John L Bixby and VanceP Lemmon 96 Well electroporation method for transfection of mammalian central neurons BioTechniques Vol 41 no 5 2006 K Regha AjitK Satapathy and Malay K Ray Re...

Page 10: ...very to Patients Molecular Therapy 17 9 1637 1642 Sep 2009 Danner S et al Seminiferous tubule transfection in vitro to define post meiotic gene regulation Reproductive Biology and Endocrinology 7 67 2009 Benton C et al Modest PGC 1_ Overexpression in Muscle in Vivo Is Sufficient to Increase Insulin Sensitivity and Palmitate Oxidation in Subsarcolemmal Not Intermyofibrillar Mitochondria The Journal...

Page 11: ...E Sowers A E and Jordan C A eds 1989 Electroporation and Electrofusion in Cell Biology Plenum Press New York 581 pp Bartoletti D C Harrison G I Weaver J C 1989 The number of molecules taken up by electroporated cells quantitative determination FEBS Lett 256 4 10 Djuzenova C S Zimmermann U Frank H Sukhorukov V L Richter E Fuhr G 1996 Effect of medium conductivity and composition on the uptake of pr...

Page 12: ... is characterized by two parameters the field strength kV cm and the time constant These parameters can be adjusted by varying voltage and capacitance settings to achieve a wide pulse gradient Field Strength The potential difference between two points electrodes in Volts divided by the distance between the electrodes called gap and expressed in cm Expressed as V cm or kV cm This is true only if th...

Page 13: ... respiration when compared to bicarbonate buffers which are also commonly used in cell culture RPMI is a form of medium used in cell culture and tissue culture It has traditionally been used for growth of Human lymphoid cells This medium contains a great deal of phosphate and is formulated for use in a 5 carbon dioxide atmosphere Opti MEM is an improved Minimal Essential Medium MEM that allows for...

Page 14: ...s are 350 watts In the USA the power cord has a standard three prong plug Installation Once you have determined that the components of the system have not sustained any obvious damage in shipment proceed with the installation The location of the BTX Gemini Twin Waveform Electroporator should be a dry level sturdy surface free from extremes in ambient temperature dust or chemical exposures Unpack t...

Page 15: ...y list Pre Pulse Resistance Measurement Used to measure the resistance of the sample prior to delivering the DC pulse Run Protocol Used to deliver the pulse protocol to the sample Scroll Down Used to scroll down in a display list Scroll Up Used to scroll up in a display list Settings Access the settings menu used to adjust the following parameters Date and Time Audible Alarms Backlighting and soft...

Page 16: ...the keypad to type the date Tap Time icon and use the keypad to type the time Tap the Green Check icon to save and return to the settings screen NOTE You may change the date and time format by tapping icons to the right of the given values Setting Audible Alarm Preferences On the home screen tap the Gears icon Tap Audible Alarms icon Select preferred audible alarms by tapping the icons to activate...

Page 17: ...ht icon Tap Device Information icon Select preferred brightness by tapping the icons indicating brightness Tap the Green Check icon to save and return to the settings screen View device information Tap the Green Check icon to return to the settings screen Software Setup Displaying Device Information On the home screen tap the Gears icon INFO BACKLIGHT BACKLIGHT KEYPAD not shown ...

Page 18: ...desired cells Preset Protocols Using Preset Protocols continued Review parameters Tap Omega icon to measure pre pulse load resistance With load measurement OK press the Circle icon to run protocol Once protocol is complete data regarding your pulse is displayed and stored in the systems logs BTX Gemini X2 only for future use You may continue pulsing go back to the protocol select screen or to the ...

Page 19: ...n Double tap desired cell type Scroll through available cells Tap desired cells Tap Folder icon Preset Protocols Customizing a Preset Protocol continued Use keyboard to name copy of protocol Tap the Green Check icon to save The message board will turn green and alert the user that the method has been copied Tap the Home icon On the home screen tap User Protocols icon KEYBOARD ...

Page 20: ... Preset Protocols Customizing a Preset Protocol continued Scroll through user protocols to locate the newly saved method Double tap the newly saved method Tap the parameter s requiring customization Proceed to modify selected parameters Tap the Green Check icon to proceed to the run screen PROTO EDIT ...

Page 21: ...ck icon to save Performing Experiments Creating New Protocols On the home screen tap User Protocols icon Creating an Exponential Decay Wave Protocol Tap the Exponential icon Tap the Green Check icon to save Double tap your newly saved protocol Tap the Voltage icon Use the numeric keypad to set voltage Tap the Green Check icon to save PROTO SET TYPE EDIT SCREEN SET VOLUME ...

Page 22: ...ng an Exponential Decay Wave Protocol continued Tap the Res Cap icon NOTE LV and HV mode capacitance values differ Use the numeric keypad to set resistance value NOTE Intervals must only be set when performing multiple pulsing NOTE Only the BTX Gemini X2 can perform multiple pulsing with Exponential Decay Wave the BTX Gemini SC cannot perform multiple pulsing with Exponential decay waves Tap the I...

Page 23: ...d Tap the Gap icon View your protocol To run tap the Omega icon Tap the Green Check icon to proceed to the run screen With load measurement OK press the Go icon to run protocol Once protocol is complete data regarding your pulse is displayed and stored in the systems logs BTX Gemini X2 only for future use You may continue pulsing go back to set up screen or to the home screen NOTE See the Protocol...

Page 24: ...he numeric keypad to set voltage Tap the Green Check icon to save Performing Experiments Creating New Protocols continued Creating a Square Wave Protocol Tap the Square Wave icon Tap the Green Check icon to save Tap the Duration icon Use the numeric keypad to set the pulse length Tap the Green Check icon to save EDIT SCREEN SET VOLTAGE EDIT SCREEN SET PULSE TIME ...

Page 25: ...ing New Protocols continued Creating a Square Wave Protocol continued Tap the Pulses Interval icon Tap the Interval sec icon Use the numeric keypad to set the pulse interval Tap the Green Check icon to save Tap the Gap icon Use the numeric keypad to set the gap between electrodes Tap the Green Check icon to save Tap the Green Check icon to proceed to the run screen EDIT SCREEN SET PROTO GAP ...

Page 26: ...tocol Once protocol is complete data regarding your pulse is displayed and stored in the systems logs BTX Gemini X2 only for future use You may continue pulsing go back to the set up screen or to the home screen Performing Experiments Creating New Protocols continued Creating a Square Wave Protocol continued View your protocol To run tap the Omega icon ...

Page 27: ...ement and DC pulses Tap the Go icon to run protocol The Square icon can be used to stop the protocol at any time NOTE Note the message box It will indicate how many pulses should be set in the HT plate handler This number will be a combination of both pre pulse resistance measurements and DC pulses NOTE Generator will display readings of the 1st and last pulse of each column Using an HT Plate Hand...

Page 28: ...iew parameters Position electrodes on area being electroporated To run tap the Omega icon With load measurement OK press the Go icon to run protocol While the pulse is being delivered the Square icon can be used to stop the protocol at any time NOTE Generator will display readings after each pulse Using an HT Plate Handler Gemini X2 only continued Once protocol is complete data regarding your puls...

Page 29: ...com 29 Using Specialty Electrodes Gemini X2 Only Once protocol is complete data regarding your pulse is displayed and stored in the systems logs for future use You may continue pulsing go back to the settings screen or the protocol select screen or to the home screen Using Speciality Protocols ...

Page 30: ...the Symbols Numbers button 123 to display the list of available numbers and symbols that can be included in the Protocol Name To switch the keyboard back to letters mode choose the Letters button ABC 15 characters maximum 5 When finished entering the new Protocol Name choose Accept to save your changes and return to the File Options screen 6 Choose BACK to return to the Mode Selection screen Renam...

Page 31: ...ne com 31 On the home screen tap the User Protocols icon Tap the protocol you wish to delete Tap Folder icon Tap Delete icon Confirm delete by tapping the Delete icon again Managing Protocols Deleting a Protocol NOTE Preset protocols cannot be deleted Protocol has been removed from the menu ...

Page 32: ...uble tap protocol you wish to password protect View the protocol Tap the Open Lock icon Managing Protocols Confirm password protection by tapping the Open Lock icon again Use the numeric keypad to set the password Tap the Green Check icon to save The protocol is now locked against modifications To Unlock press the lock icon Type in the password then Tap the Green Check icon LOCK ...

Page 33: ...r to the PC using a USB cable 2 Turn generator on 3 Start Protocol Manager 4 Select the proper COM port Overview Protocol Manager allows the user to Upload protocols from the Gemini X2 generator to a PC Download protocols from PC to the Gemini X2 generator Upload log files from the Gemini X2 generator to PC Log File Control PC to Generator Protocol Control Generator to PC Protocol Control Protocol...

Page 34: ... Download PC to Generator 1 Click Choose Protocol 2 Select Protocol s to download 3 Click Open 4 Protocol name being downloaded is shown Protocol Manager Software NOTE If a protocol already exists on the Gemini X2 generator Protocol Manager will not overwrite the file if you attempt to re download it ...

Page 35: ...your desired files or folders and then click Transfer 3 Select the drive or folder location Troubleshooting Protocol Manager 1 Do not upload download Protocols while generator is running Program works best while generator is idle on Protocol setup screen 2 Order of Operations A Connect USB B Turn generator on C Start Protocol Manager program NOTE Log Files are saved as a tab delimited text file Th...

Page 36: ...provide a location to install your application files There is a known issue with this installer where it will not actually create this directory Overview Remote Screen Interface allows you to control the Gemini remotely When you click on the computer screen you control the generator While the Gemini generator is connected to the computer via USB open the Control Panel and select System and Securit...

Page 37: ...10 3000 10 V steps Voltage Accuracy LV Mode 5 5 HV Mode 5 5 Time Constant Range LV Mode 1 ms to 5 158 s 1 25 ms to 3 275 s 1 25 ms HV Mode 0 5 ms to 133 875 ms 0 5 ms to 50 ms 0 5 ms Multiple Pulsing Multiple Pulsing 1 to 2 depending on device s current amplitude limit No Pulse Interval 5 to 30 s N A Capacitance in µF LV Mode 25 3275 µF in 25 µF steps 25 3275 µF in 25 µF steps HV Mode 10 25 35 50 ...

Page 38: ...rking CB Scheme EuRoHS Yes Yes Dimensions H x W x D inches 12 75 x 11 25 x 8 5 12 75 x 11 25 x 8 5 Weight 16 lb 16 lb Warranty 2 years 1 year Atmospheric Specifications Operating Temperature 4 C to 40 C 40 F to 104 F Storage Temperature 10 C to 70 C 14 F to 158 F Operating Humidity See Chart Below Storage Humidity 20 to 80 RH non condensing Mode of Operation Continuous Classification Class I Pollu...

Page 39: ...Make sure the power cord is disconnected from the main supply before servicing the fuse 2 Use only Type 3AG 1 4 x 11 4 in 5 amp 250 volt Type T time delay fuses Turn off power and remove power cord from power module Use a straight blade screw driver to pry open the access door Remove the fuse holder and then remove the fuses from this holder as shown in figure below Replace fuses and then replace ...

Page 40: ...ual CommPort Driver supplied with the BTX Gemini X2 1 Connect the Gemini X2 generator to the computer via USB Open the control panel and select System and Security and then System Click Device Manager on the left hand menu Pin Signal 1 5V 2 Data 3 Data 4 GND Mating Face Upgrading Gemini Series Software 2 Two Flash update applications will be displayed in the device manager under Other Devices Righ...

Page 41: ...essage regarding the driver publisher is displayed choose Install this driver software anyway USB Virtual Commport Driver Installation continued 4 Click Browse and navigate to the generator drivers folder located on the CD supplied with the generator Select the file BTX USB CDC inf Make sure the Include Subfolders box is checked and then click Next Upgrading Gemini Series Software ...

Page 42: ...xit boot loader mode without upgrading the software power cycle the generator 4 From the CD provided with the Gemini X2 generator open the Device Updater Application Upgrading Gemini Series Software NOTE Prior to upgrading users will need to install USD drivers as well as the bootloader driver file Reference Virtual Comport Driver Installation 5 Click Open Update File and browse to the BTX Gemini ...

Page 43: ...onditions 1 Sample resistance is very low for the voltage selected 2 Gap size is too small for the voltage selected 3 Pulse duration is too long No_HB_from_GUI Communication error Hardware error Please contact the manufacturer Sample Resistance Out Of Range The measured sample load is out of range Increase the sample resistance This can be achived by 1 Increasing the gap size 2 Reducing the volume...

Page 44: ...Failure Notification If power is interrupted during operation an alarm will sound to alert the user to the interruption When power is restored an onscreen message is displayed to indicate the interruption Out of Range A value was entered or encountered in a protocol that was beyond the generator s limits NOTE It is recommended that you save your protocols periodically by uploading them to a PC ...

Page 45: ...ers Verify delivery of the pulse and pulse parameters Is the voltage correct Chamber gap Pulse length or appropriate instrument settings Number of pulses If so follow Optimization Guidelines outlined Low Viability Verify physical biological chemical parameters Is the voltage chamber gap pulse length time constant pulse number and other instrument settings correct If so reduce voltage pulse length ...

Page 46: ... 96 Well Plate 2 mm gap 125 µl 1 ea 45 0452 96 Well Plate 4 mm gap 250 µl 1 ea Reagents 45 0802 BTXpress 5 ml 1 ea 45 0803 BTXpress 5 ml with 2 mm gap Cuvettes pk 50 1 ea 45 0804 BTXpress 5 ml with 4 mm gap Cuvettes pk 20 1 ea 45 0805 BTXpress 10 ml 1 ea 45 0806 BTXpress 10 ml with 2 mm gap Cuvettes pk 50 1 ea 45 0807 BTXpress 10 ml with 4 mm gap Cuvettes pk 40 1 ea 47 0002 Cytoporation Media T 50...

Page 47: ... needs 45 0216 1 Ea 45 0507 Petri Dish Tissue Chamber 15 x 15 mm Kit 1 Ea 45 0100 Petri Dish Electrode 2 mm gap 90 mm Well Dish Kit 1 Ea 45 0130 Petri Pulser for 6 Well Plates 35 mm Well Kit 1 Ea 45 0490 Tissue Slice Chamber 7 x 7 mm Kit 1 Ea Order No Description UOM 45 0491 Tissue Slice Chamber 7 x 7 mm Dish needs 45 0492 45 0503 45 0204 1 Ea 45 0492 Tissue Slice Wand 7 mm needs 45 0491 45 0503 4...

Page 48: ...Holliston Massachusetts USA 01746 1388 using our toll free number 1 800 272 2775 US Only or 508 893 8999 E mail techsupport btx harvardapparatus com Goods will not be accepted for return unless an RMA Returned Materials Authorization number has been issued by our customer service department The customer is responsible for shipping charges Please allow a reasonable period of time for completion of ...

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