4D-Nucleofector™ Manual –
Bioscience Solutions
39
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Page 1: ...Bioscience Solutions 4D Nucleofector System Manual For Research Use Only...
Page 2: ...System conveys to the buyer the non transferable right to use the system as well as Lonza s proprietary Nucleofector Technology for research conducted by the buyer whether the buyer is an academic or...
Page 3: ...ofector Y Unit 18 2 10 1 Defining a New Experiment 18 2 10 2 Loading Samples 19 2 10 3 Running the Experiment 19 2 11 4D Nucleofector LV Unit 20 2 11 1 Using the 1 mL Nucleocuvette Cartridge Fixed Vol...
Page 4: ...owing reliable transfection of adherent cells and cells in suspension Transfection of Any Substrate Nucleofector Technology offers high flexibility within applications since the same transfection para...
Page 5: ...ioscience Solutions 5 The 4D Nucleofector System Support Teams Europe Scientific Support 32 87 321 611 scientific support eu lonza com North America Scientific Support 1 800 521 0390 toll free scienti...
Page 6: ...power supply before cleaning and disinfecting the case Useadampclothtowipedowntheoutercasewithwateror70 80 ethanol Do not use any aerosols for cleaning Avoid wetting the cuvette holder within the cuv...
Page 7: ...nit 1 mL or LV Nucleocuvette Cartridge for LV Unit 24 well Dipping Electrode Array forYUnit UsingconsumablesfromanyothersourcethanLonzawill preclude all warranty and liability claims Do not alter the...
Page 8: ...mple rescue also see 3 2 using wheel accessible after removal of side panel via screw behind termination plug A typical 4D Nucleofector System includes 1 One 4D Nucleofector Core Unit 2 At least one f...
Page 9: ...for completeness 2 Stack the units with the Core Unit containing the touch screen on top figure 2 4 3 Connect the units figure 2 5 by using the interface cables Connect the interface outlet port of t...
Page 10: ...ector System Core Unit and functional units Figure 2 6 Main screen 2 7 2 Adjusting the Position of the Touch Screen The touch screen of the 4D Nucleofector System can be set at four angles 0 30 45 and...
Page 11: ...onfirm selection or execute a program A Z Z A Top 10 Last 10 Sort a list alphabetically display the most frequent 10 items or the most recent 10 items Magnifier symbol Activate search functions i Disp...
Page 12: ...s Load Samples chapter 2 9 2 10 or 2 11 Execute Nucleofection Experiment chapter 2 9 2 10 or 2 11 Prepare Samples According to Cell type Specific Optimized Protocol 2 8 General Instructions for Runnin...
Page 13: ...ntaining cells and substrate The selected Nucleofection program will be applied to this position 2 No DNA Negative control Nucleofection program applied to vessel with cells but without substrate 3 No...
Page 14: ...line of the list The experiment selected will be highlighted Confirm your selection by pressing OK 5 By pressing on the icons displaying the Nucleofection vessels or wells you can check the settings...
Page 15: ...Z to find conditions more quickly a Select the desired cell type code by tapping on the appropriate line of the cell list The cell type selected will be highlighted For additional information about t...
Page 16: ...leocuvette Strip theyellowpinattherearend of thestripmustbevisible figure2 14 C Ifthestripismounted inthewrongorientationitsrearendwillstayabovethestripholder and the yellow pin is hardly visible figu...
Page 17: ...te s Proceed willbe displayed figure 2 15 C Load next samples and press YES to continue or press NO to interrupt the experiment 12 When the experiment is complete a result file summarizing the Nucleof...
Page 18: ...line of the cell list The cell type selected will be highlighted For additional information about the cell type selected press i b To confirm your selection press OK c If required modify pulse code b...
Page 19: ...the softwarewillcheckforusedwellsandofferyoudifferentoptionshow to continue 2 10 3 Running the Experiment 12 Afterloadingthesamplespress START toruntheexperiment figure 2 18 A 13 The progress of the...
Page 20: ...part area 3 Injection port rear side 4 Air outlet port with 0 2 M sterile filter Components of the LV Nucleocuvette Cartridge figure 2 19 B C 1 Cartridge 2 Venting tube with 0 2 M sterile filter 3 Inl...
Page 21: ...of defining solution and program code via the CELL TYPE PROGRAM both parameters can also be selected manually e g in casenopredefinedcelltypeprogramisavailable Foraddingnewcell type programs please r...
Page 22: ...y a color code green for OK figure 2 22 C yellow in case of few errors and red in case of multiple errors figure 2 22 D If errors occurred you can check for more details by pressing More details There...
Page 23: ...KIP and continue with step 14 4 Upon pressing YES the pinch valves will automatically open for 30 secondsallowingtoconvenientlyinsertthetubesintothethreepinch valves and a new window will appear figur...
Page 24: ...o cell type specific protocol and mount it into a 4D Nucleofector LV Reservoir Rack figure 2 23 I 12 RemovetheredcapfromtheSpirosconnectorontheoutlettubeofthe cartridge and connect it to the swabbable...
Page 25: ...apter 2 12 3 17 Optional At this stage or at step 20 you can save your defined experimentforfutureusebypressingthe SAVE button figure2 24 D Akeyboardwillappearallowingyoutodefineaname max length 26 ch...
Page 26: ...pressing START H I Figure 2 25 Setup Checklist for LV Nucleocuvette Cartridge 1 Cartridge inserted in slot 2 Short venting tube with filter attached to the male Luer connector on the front plate 3 Tub...
Page 27: ...rs figure 2 26 E and red in case of multiple errors figure 2 26 G If errors occurred you can check for more details by pressing More details The result details figure 2 26 F H will display the type of...
Page 28: ...ttings list Firmware update update the system software see chapter 2 12 7for details Device cleaning opens the tray of the functional modules to remove it for cleaning see chapter 2 12 5 for details S...
Page 29: ...he NEW field Edit existing programs by pressing EDIT or save your programs onto a USB stick by pressing SAVE figure 2 27 E To create a new custom program or to edit an existing program Press NEW or se...
Page 30: ...D Nucleofector SystemwiththePCEditorsoftware seechapter2 13 Itisnecessaryto perform synchronization with each firmware update To start synchronization Insert a USB stick into the USB port on the front...
Page 31: ...drive Insert 4D Nucleofector user information USB stick into the USB port of your PC Copy all files from the 4D Nucleofector PC Editor software folder to your hard disk preferably into a folder calle...
Page 32: ...tion figure 2 29 B Select the USB stick and press SAVE Unplug the USB stick from your PC as soon as the experiment file has been saved NOTE Do not unplug the stick while saving Turn on the 4D Nucleofe...
Page 33: ...easing DNA amount may lead to higher transfection efficiency but at the same time result in higher cell mortality Cell number in Nucleofection sample too high or too low Please use the cell numbers re...
Page 34: ...irection of the red arrow as far as it will go figure 3 1 B Model variant B Newer Unplug system from main power Remove the terminator cap or cable connection to next unit figure 3 1 C Insert screw dri...
Page 35: ...argecircumventingthesampleandis oftenaccompaniedbyaflashandanoise Thisproblemisusuallycaused by imperfect cuvettes or cuvette filling The 4D Nucleofector System is equipped with a hardware safety feat...
Page 36: ...e of dipping electrodes that had been used before Err 22 Used well A weak pulse occurred Efficiency may be sub optimal Re used dipping electrode or inappropriate Nucleofector Solution Clear the error...
Page 37: ...a for the Licensed Process and or the Purchased Device is complete Lonza does not warrant either that the use of the License does not infringe third parties rights or does not cause damages to third p...
Page 38: ...n interfaces interface RS422 USB Lonza interface for module connection Shuttle HV Out X Unit only Spannungsbegrenzung voltage limitation limitation de tension 1500 V Strombegrenzung current limitation...
Page 39: ...4D Nucleofector Manual Bioscience Solutions 39 6...
Page 40: ...partyowners Theinformationcontainedhereinisbelievedtobecorrect and corresponds to the latest state of scientific and technical knowledge However nowarrantyismade eitherexpressedorimplied regarding it...
