Lonza 4D-Nucleofector Core Unit Manual
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Summary of Contents for Lonza 4D-Nucleofector Core Unit
- Page 1 Bioscience Solutions 4D-Nucleofector™ System Manual For Research Use Only...
- Page 2 The purchase of the 4D-Nucleofector™ 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 for-profit entity). For further details about the license please refer to chapter 5.
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Page 3: Table Of Contents
2.12 Additional Settings and Options 2.12.1 Experiments 2.12.2 Results 2.12.3 Custom Programs 2.12.4 Synchronize 2.12.5 Cleaning the Nucleofection Vessel Tray (X or Y Unit) 2.12.6 Lonza Programs 2.12.7 Firmware Update 2.13 PC Editor Software 2.14 96-well Shuttle™ Mode Troubleshooting 4D-Nucleofector™ Manual – Bioscience Solutions... -
Page 4: The Nucleofector™ Technology
Since its introduction in 2001, Nucleofector™ Technology has transformed transfection. Nucleofection has proven to be a reliable and reproducible At Lonza, we believe that listening to our customers is the best way easy-to-use technology, suited to a wide variety of applications. An ever to develop innovative products that meet your needs. - Page 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) scientific.support@lonza.com 4D-Nucleofector™ Manual – Bioscience Solutions...
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Page 6: Operating Instructions
5. Certain commercial application is inside a receptacle incorporated in the power socket of the Core Unit allowed under Lonza’s license for for-profit-entities, as set forth in the (see figure 2.0). Blown fuses can easily be replaced. Disconnect the license. -
Page 7: Waste Disposal
– Do not plug any non-Lonza-certified connectors into the external high Lonza disclaims all warranties and shall in no event be liable for any kind voltage electrode sockets at the rear side of the 4D-Nucleofector™ of damages caused by or arising out of any operation or use in violation System. -
Page 8: Nucleofector™ System Components 8 5 Purchaser Notification
2.5 4D-Nucleofector™ System Figure 2.1: 4D-Nucleofector™ System before assembly Components By definition the 4D-Nucleofector™ System is a modular system offering maximum flexibility. Therefore, a complete system may vary in the number and type of components it contains. Furthermore, a system may consist of minor model variants of the units. -
Page 9: Set-Up Instructions
15. Lonza interface inlet port to connect functional units 2. Stack the units with the Core Unit (containing the touch screen) on 16. Lonza interface inlet port of last functional unit with top (figure 2.4) termination plug 3. Connect the units (figure 2.5) by using the interface cables: 17. -
Page 10: General Use Instructions 10 6 Technical Specifications
Details of the Nucleofection process can be found in the cell- angles (0°, 30°, 45°, and 60°), providing convenient access for the user. type specific Optimized Protocols (www.lonza.com/optimized-protocols). Press the label on top of the display frame to unlock the display from its home position. -
Page 11: Operating Software
2.7.3 Operating Software The 4D-Nucleofector™ System is operated via a 5.7 inch touch screen display and controlled by intuitive software. Table 2.2 will familiarize you with the functions of the touch fields and the basic features of the Figure 2.8: Software screen elements operating software. -
Page 12: General Instructions For Running A Nucleofection Experiment
Select Vessel Type (chapter 2.8.2) Load Predefined Experiment Define New Experiment (chapter 2.8.4) (chapters 2.9, 2.10 or 2.11) Lonza Template Saved User Experiment Use As Is Modify Prepare Samples According to Cell-type Specific Optimized Protocol Load Samples (chapter 2.9, 2.10 or 2.11) Execute Nucleofection Experiment (chapter 2.9, 2.10 or 2.11) -
Page 13: Unit And Vessel Selection
The “Cell Type Program” list can be extended by the user with customized cell-type codes (e.g., after a cell line optimization; 2.12.3). Lonza defined programs are highlighted in blue while custom program codes will be highlighted in black. Solution In case the recommended Nucleofector™... -
Page 14: Using A Predefined Experiment
Figure 2.11: Using a predefined experiment (example: X Unit) A predefined experiment can either be a ready-to-use template pre- installed by Lonza (for X and Y Unit only) or a previous experiment that was saved by the user. The use of predefined experiments is described showing the X Unit as an example, but the procedure would be the same for Y or LV Unit. -
Page 15: Nucleofector™ X Unit
2.9 4D-Nucleofector™ X Unit Figure 2.12: Vessel types for 4D-Nucleofector™ X Unit The 4D-Nucleofector™ X Unit can handle two types of vessels: – 16-well 20 µL Nucleocuvette™ Strips for cell numbers from 2 x 10 1 x 10 cells – Single 100 µL Nucleocuvette ™ Vessels for cell numbers from 2 x 10 to 2 x 10 cells 2.9.1... -
Page 16: Loading Samples
– For the 100 µL Nucleocuvette™ Vessel, the cuvettes should be C Right orientation D Wrong orientation mounted such that the label LONZA on the lid is readable and the opening clip is at the front (figure 2.14, G, H). Bioscience Solutions – 4D-Nucleofector™ Manual... -
Page 17: Running The Experiment
2.9.3 Running the Experiment Figure 2.15: Experiment progress (X Unit) 10. After loading the samples press “START” to run the experiment (figure 2.15, A). 11. The progress of the experiment is indicated by changing the color of the cuvette or well positions (figure 2.15, B) (for color codes see chapter 4). -
Page 18: Nucleofector™ Y Unit
2.10 4D-Nucleofector™ Y Unit Figure 2.16: Experiment definition (Y Unit) 2.10.1 Defining a New Experiment 1. After unit selection (see chapter 2.8.2) a screen appears that allows you to define a new experiment or select a previously saved experiment by pressing “Choose existing experiment” (figure 2.16, A). 2. -
Page 19: Loading Samples
2.10.2 Loading Samples Figure 2.17: Loading of samples (Y Unit) 9. Insert the 24-well Dipping Electrode Array into the 24-well culture plate containing your Nucleofection samples. Make sure that the dipping electrode array is inserted in the right orientation. 10. Place 24-well plate with inserted dipping electrode array into the retainer of the 4D-Nucleofector™... -
Page 20: Nucleofector™ Lv Unit
2.11 4D-Nucleofector™ LV Unit Figure 2.19: Nucleocuvette™ Cartridge types for LV Unit The 4D-Nucleofector™ LV Unit can handle two types of vessels, the fixed volume 1 mL Nucleocuvette™ Cartridge for cell numbers up to 1x10 cells and the scalable LV Nucleocuvette™ Cartridge for cell numbers up to 1x10 cells. -
Page 21: Using The 1 Ml Nucleocuvette™ Cartridge
2.11.1 Using the 1 mL Nucleocuvette™ Cartridge Figure 2.20: Experiment definition (LV Unit, 1 mL Nucleocu- vette™ Cartridge) (Fixed Volume) 2.11.1.1 Defining a New Experiment 1. After unit and vessel type selection (see chapter 2.8.2) a screen appears which allows you to define experiment parameters select a previously saved experiment by pressing “Choose existing experiment”... -
Page 22: Running The Experiment
2.11.1.3 Running the Experiment Figure 2.22: Experiment progress (LV Unit, 1 mL Nucleocuvette™ Cartridge) 11. When the cartridge is inserted, press “START” to initiate the Nucleofection process (figure 2.22, A). 12. The progress of the experiment is indicated by a progress bar (figure 2.22, B). -
Page 23: Using The Lv Nucleocuvette™ Cartridge (Scalable Volume)
Figure 2.23: Mounting the LV Nucleocuvette™ Cartridge 2.11.2 Using the LV Nucleocuvette™ Cartridge (Scalable Volume) Before starting to prepare any cells – Insert LV Nucleocuvette™ Cartridge including all tubes into the 4D-Nucleofector™ LV Unit (see steps 1-10 below) – Connect outlet reservoir (see steps 11-12 below) –... - Page 24 9. Insert the upper tube into the upper pump and close the flap (figure Figure 2.23: Continued 2.23, H). 10. Repeat with the lower tube and the lower peristaltic pump (optional if working with 2 reservoirs). 11. Fill outlet reservoir with medium (for detailed recommendations, please refer to cell type-specific protocol) and mount it into a 4D-Nucleofector™...
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Page 25: Defining A New Experiment
2.11.2.2 Defining a New Experiment Figure 2.24: Experiment definition (LV Unit, LV Nucleocuvette™ Cartridge) 14. Press “NEXT” (figure 2.24, A) to define experiment parameters or select a previously saved experiment by pressing “Choose existing experiment” (figure 2.24, B). 15. Press the field “CELL TYPE PROGRAM” to choose predefined Nucleofection conditions from a cell type list (figure 2.24, C). - Page 26 Once samples have been prepared finalize experiment definition: Figure 2.24: Continued 26. Continue with entering the volume(s) that you are going to process (figure 2.24, H). a. If you work with one reservoir (or bag) containing a premixed suspension of cells and substrate, enter the total volume of the cell suspension you want to process.
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Page 27: Running The Experiment
2.11.2.3 Running the Experiment Figure 2.26: Experiment definition (LV Unit, LV Nucleocuvette™ Cartridge) 28. Check defined experiment including volumes again and press “START” to initiate the Nucleofection process (figure 2.26, A). 29. Before starting the system performs a calibration step (figure 2.26, 30. -
Page 28: Additional Settings And Options
USB port in front of the Core Unit (see chapter 2.12.2 for more information) – Lonza programs – version information about the cell type list and option to upload a new cell type program list (see chapter 2.12.6) –... -
Page 29: Results
“Cell Type Program” list and can be differentiated from Lonza defined code by color: Blue: Lonza defined cell type program codes Black: Customer defined codes. You can create new custom programs by pressing the “NEW” field. Edit existing programs by pressing “EDIT” or save your programs onto a USB stick by pressing “SAVE”... -
Page 30: Synchronize
NOTE: More comprehensive firmware updates may not be possible via – Open the “Settings” menu USB upload and require return of the system to Lonza. – Press “Device Cleaning”. – The drawer will open, giving access to the Nucleofection vessel tray. -
Page 31: Pc Editor Software
2.13 PC Editor Software Figure 2.28: Files comprising the 4D-Nucleofector™ PC Editor The PC Editor is a PC-compatible version of the 4D-Nucleofector™ User Interface. It can be used to define experiments on the PC and then transfer them to the 4D-Nucleofector™ System via USB stick. This chapter describes the installation of the 4D-Nucleofector™... - Page 32 Transfer of Experiments from the PC Editor Figure 2.29: Transferring experiments from the PC Editor – Open PC Editor by double clicking on the file 4D-Nucleofector_Editor.exe. – Define and save your experiment with the PC Editor – Plug USB stick into a USB port of your PC. –...
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Page 33: Suboptimal Transfection Results
2.14 96-well Shuttle™ Mode Figure 2.30: Shuttle connectivity on the 4D-Nucleofector™ System To operate the 96-well Shuttle™ Device a 4D-Nucleofector™ System comprising Core and X Unit is required (for details about operating the “Nucleofector™ 96-well Shuttle™ System” please refer to the respective 96-well Shuttle™... -
Page 34: Rescue Of Samples (X And Y Unit)
– Open the front flap manually (figure 3.1, E) – Pull the tray cautiously towards you (figure 3.1, F) – Remove the Nucleocuvette™ Vessels and rescue your cells – Contact your Lonza representative to solve potential technical issue Bioscience Solutions – 4D-Nucleofector™ Manual... -
Page 35: Nucleofector™ System
Arcing is a complete or partial discharge circumventing the sample and is often accompanied by a flash and a noise. This problem is usually caused If the problem cannot be resolved, please call Lonza’s Scientific by imperfect cuvettes or cuvette filling. The 4D-Nucleofector™ System is... - Page 36 Error Code What Happened? Possible Error Procedure Err1A No pulse generated Internal communication failure, internal Clear the error message by pressing any button and try to apply error or device possibly defective. program a second time. If Err1A re-occurs, switch off the device, check the interface cable, wait for 2 seconds and switch on again.
- Page 37 Purchased Device or Licensed Process provided to Buyer by Lonza, or (c) the gross negligence or willful misconduct of Buyer or any Lonza represents and warrants that, it is the holder of intellectual property, including patents, of its affiliates in performing any activity relating to the subject matter hereof.
- Page 38 Y Unit: 8-10 reactions (using Dipping Electrodes) LV Unit: 2-3 mL (using LV Nucleocuvette™ Cartridges) Schnittstellen / interfaces / interface RS422 Lonza interface for module connection Shuttle HV Out (X Unit only) Spannungsbegrenzung / voltage limitation / limitation de tension 1500 V...
- Page 39 4D-Nucleofector™ Manual – Bioscience Solutions...
- Page 40 The Nucleofector™ Technology is covered by patent and/or patent pending rights owned by the Lonza Group Ltd or its affiliates. Unless otherwise noted, all trademarks herein are marks of the Lonza Group or its affiliates. Nucleofector™ Kits contain a proprietary nucleic acid coding for a proprietary copepod protein fluorescent protein intended to be used as a positive control with Lonza products only.
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