Lonza 4D-Nucleofector Manual
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Summary of Contents for Lonza 4D-Nucleofector
- Page 1 Enabling a Healthier World 4D-Nucleofector ® Manual Your guide to setup and usage of the 4D-Nucleofector System ® 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 propri- etary Nucleofector Technology for research ® conducted by the buyer (whether the buyer is an academic or for-profit entity). For further de- tails about the license please refer to chapter 5.
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Page 3: Table Of Contents
Consignes de sécurité - à lire attentivement Waste disposal Suboptimal transfection results 4D-Nucleofector System components ® Rescue of samples (X and Y Unit) Set-up instructions 2.7.1 Turning on the 4D-Nucleofector System ® Error Codes of the 2.7.2 Operating software ® 2.7.3 Turning off the 4D-Nucleofector... -
Page 4: Nucleofector Technology
® the process, please refer to our cell type-specific Optimized With over 325 protocols optimized by Lonza’s R&D Team for Protocols in our cell database. cell lines and primary cells and more than 750 entries in the... -
Page 5: Moving Transfection Into The Fourth Dimension
Nucleofector Platform: ® fourth dimension At Lonza, we believe that listening to our customers is the • A novel conductive polymer based cuvette system com- best way to develop innovative products that meet your pletely free of toxic metal ions needs. -
Page 6: Operating Instructions
The fuse receptacles (each marked with an arrow) in humans. are then also levered open using the screwdriver. License statement Lonza is holder of various patents, patent applications, cop- yrights and technical and scientific experience with respect to Nucleofector Technology. Use of Lonza’s Nucleofector ®... -
Page 7: Consignes De Sécurité - À Lire Attentivement
It will destroy the device. • Lonza disclaims all warranties and shall in no event be • Only use the power cord that comes with the device. liable for any kind of damages caused by or arising out •... -
Page 8: Waste Disposal
Lonza Cologne décline toute garantie et ne pourra en au- d‘explosion, ni pour une utilisation avec des milieux in- cun cas être tenue responsable de tout type de dommages flammables ou explosifs. -
Page 9: Nucleofector ® System Components
8. Lonza interface inlet port to connect functional units 9. Lonza interface inlet port of last functional unit 10. Lonza interface cable connected to outlet and inlet port 11. Lonza interface outlet port of functional unit 12. Lonza outlet port of last functional unit with termination... -
Page 10: Set-Up Instructions
2.7 General use instructions edit existing cell type specific protocols Time and date (2) Shows time and date (changes can be made in This section gives an overview of the 4D-Nucleofector ® the Settings menu) System operating software. Details of the Nucleofection ®... -
Page 11: Turning Off The 4D-Nucleofector ® System
Nucleofection Experiment independ- ® By selecting the vessel type a new screen opens where ent of the different 4D-Nucleofector Units. The detailed ® either a new experiment can be started or one of the pre- handling of each unit is described in subsequent chapters defined experiments can be loaded. -
Page 12: Experiment Parameters - Overview
Via the “Cell type code” a predefined combina- tion of a “Pulse code” and a “Solution” can be selected. All Lonza optimized conditions for a cell type are available in the “Cell type code” drop down menu. Customized cell type codes... -
Page 13: Using A Predefined Experiment
A predefined experiment can chapters 2.9, 2.10, 2.11 or 2.12) once loaded. You can either be a ready-to-use template pre-installed by Lonza (i.e. save the changed experiments under a new name by pressing “Save”. Alternatively, you are prompted to... -
Page 14: Nucleofector® X Unit
(see chapter 2.8.4). Tap on the checkmark button to get to the Experiment Setup screen (Figure 2.15). Figure 2.13: 4D-Nucleofector Core and X Unit (A) and Nucleocuvette Ves- ®... - Page 15 ® 2.15, B). A list with all available Cell type codes (defined by Lonza or custom) will appear. Select the code that shall be used by scrolling through the menu or start typing in the search field (Figure 2.15, C).
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Page 16: Loading Samples
Vessel, the cuvettes should be mounted such that ® the label Lonza on the lid is readable and the opening clip is at the front (Figure 2.17, G, H). Figure 2.18: Experiment progress (X Unit) 1. After loading the samples, press “Start” to run the experi- ment (Figure 2.16). -
Page 17: Nucleofector Y Unit
2.15, B). A list with all available Cell type codes (defined If you want to create a new Experiment, you may tap in the by Lonza or custom) will appear. Select the code that field “Experiment name” and enter a specific experiment... -
Page 18: Loading Samples
2. Place the 24-well plate with inserted dipping elec- trode array into the retainer of the 4D-Nucleofector ® Y Unit. Well “A1” must be in the upper left position (Figure 2.22). -
Page 19: Nucleofector 96-Well Unit
By tapping on a well position one sample/well is selected, marked by a blue circle around the well position. Multiple samples/wells can be selected by dragging across the wells. Figure 2.24: 4D-Nucleofector Core Unit and 96-well Unit (A). 96-well ® Nucleocuvette Plate (B). - Page 20 ® 2.15). A list with all available Cell type codes (defined by Lonza or custom) will appear. Select the code that shall be used by scrolling through the menu or by starting to type in the search field. Enter the cell name in the field Cell type code.
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Page 21: Loading Samples
4. Press “Start” to start the same experiment again summarizing the Nucleofection Process will be ® or “Back” to return to the “Set up Experiment” displayed (Figure 2.29). screen and define a new experiment. Figure 2.29: Example of a result file. 4D-Nucleofector Manual | 21 ®... -
Page 22: Nucleofector ® Lv Unit
5. Outlet port (spinning Spiros with red dust protection cap) 6. Reservoirs (swabbable injection port with male Luer cap) NOTE: Please do not alter or disassemble the pre-assembled cartridge or reservoirs. Components of the 4D-Nucleofector LV Reservoir ® (Figure 2.31, C) 1. -
Page 23: Using The 1 Ml Nucleocuvette ® Cartridge (Fixed Volume)
1. Press the field “Cell Type Code” to choose predefined Nucleofection Conditions from a Cell type list (Figure 2.33). ® A list with all available Cell type codes (defined by Lonza or custom) will appear. 4D-Nucleofector Manual | 23 ®... -
Page 24: Loading Samples
7. A summary of the defined settings will be displayed. b. Yellow cross in case of few errors (Figure 2.36, B) Before loading samples and pressing “Start” (see c. Red dash in case of multiple errors (not shown). chapter 2.12.13), please double-check for accuracy. 24 | 4D-Nucleofector Manual ®... -
Page 25: Using The Lv Nucleocuvette ® Cartridge (Scalable Volume)
® Step 6: Verify volume settings and execute experiment (see 2.12.2.4) NOTE: In case you use bags and not Lonza LV Reservoirs for sample input and/or collection, please contact Lonza Scientific Support for maximum tube volumes. Figure 2.36: Result screens for LV Unit. (A/B) Result screen for no errors in experiment. -
Page 26: Defining A New Experiment
(>7 mm). A message will appear and the user can decide whether the process by Lonza or custom) will appear. Select the code that shall be used by scrolling through the menu or start shall be continued or aborted. In case “continue” is typing in the search field . -
Page 27: Mounting An Lv Nucleocuvette ® Cartridge
9. Repeat with the lower tube and the lower peristaltic pump (optional, if working with 2 input reservoirs). 10. Fill the output reservoir with medium (for detailed recommendations, please refer to cell type-specific protocol) and mount it into a 4D-Nucleofector ® Reservoir Rack (Figure 2.37, H). 4D-Nucleofector Manual | 27 ®... - Page 28 Return to TOC Figure 2.37: Mounting the 4D-Nucleofector LV Nucleocuvette Cartridge. ® ® (A) LV Cartridge inserted into slot and venting tube attached. (B) Setup screen for LV Cartridge. (C) Remaining time for tube insertion is shown. If the time is not sufficient, you can start the process again after the time ran up.
- Page 29 Input reservoir for substrate (optional) c. Output reservoir Figure 2.38: Setup Checklist for LV Nucleocuvette Cartridge. The picture ® reflects the GMP-grade TheraPEAK Consumables, research-grade versions ® may slightly differ but the mounting procedure stays the same. 4D-Nucleofector Manual | 29 ®...
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Page 30: Running The Experiment
11. You can repeat the same experiment by pressing monitoring. As such, it is dependent on the tubing diameter “Restart”. which may slightly vary due to raw material variations and/or tubing tension. 30 | 4D-Nucleofector Manual ®... -
Page 31: Additional Settings And Options
• Network: Shows available network connections • Version: Displays software and firmware versions of all connected 4D-Nucleofector Units. ® • Fill mode (only available if LV Unit is connected): The “Fill mode” can be adapted when using the flow-through cartridge version, the LV Nucleocuvette Cartridge. -
Page 32: Cleaning The Nucleofection ® Vessel Tray (X, Y, Or 96-Well Unit)
2.13.1.3 Version Tap on “Version“ to find out more information about the software and the Lonza program list version. The table “De- vice Versions“ displays the firmware version and the serial number of all functional units. See „2.13.4 Firmware Update“... -
Page 33: Results
Custom Cell type codes are included in alpha- an existing optimized protocol. betical order and can be distinguished from Lonza defined • Select the functional units for which the code should ap- Pulse codes by color: ply (either X, 96-well and LV Unit, or Y Unit). -
Page 34: Lonza Programs
To make sure if an upgrade is advised, please reach out to Scientific Support. Figure 2.47: (A) Message upon insertion of USB stick with respective Lonza program file. (B) message after tapping on the program update button. • Choose “Program updates available” by tapping in the field. -
Page 35: Transfer Of Experiments From The Pc Editor
– preferably into a folder called “4D-Nucleofector PC Editor” ® • Open the folder, search the file called “4D-Nucleofector ® “Editor” and double click on it to start the software. • A window will appear displaying the graphical user inter- face of the 4D-Nucleofector PC Editor. -
Page 36: Troubleshooting
To avoid a deep discharge of the battery and loss of data, switch on the system at least once a month. If the problem cannot be resolved, please call Lonza’s Scientific Support Team... -
Page 37: Rescue Of Samples (X And Y Unit)
Figure 3.1: (A/B) Rescue of samples Further steps • Manually open the front flap. • Carefully pull the tray./ • Remove the Nucleocuvette Vessels and rescue your ® cells. • Contact your Lonza representative to solve potential technical issues. 4D-Nucleofector Manual | 37 ®... -
Page 38: Error Codes Of The 4D-Nucleofector
® with a hardware safety feature that immediately detects by clicking on the well in the result file. arc formation and protects the cells from damage. After the arc interruption the 4D-Nucleofector System re- ® 4.2 Result rating (LV Unit) sumes program execution. -
Page 39: Nucleofector ® System Error Codes
Return to TOC 4.4 4D-Nucleofector System Error Codes ® The 4D-Nucleofector System detects and reports different ® types of errors. Some are just error messages, others might be indicated by a code. The table below explains the result codes (errors or results) that can be indicated in the next generation 4D-Nucleofec- System. - Page 40 User confirmed to skip Well was skipped the well as it had been used before www.lonza.com/nucleofection-error-codes If a problem cannot be resolved, please call Lonza’s Scientific Support Team. Europe Phone: + 49 221 99199 400 E-mail: scientific.support.eu@lonza.com North America Phone: 800 521 0390 (toll free) E-mail: scientific.support@lonza.com...
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Page 41: Purchaser Notification
The License is limited to research use only. The License does not include the right to use the Purchased Device, its Lonza does not warrant either that the use of the License components or materials, or the Licensed Process for any does not infringe third parties’... - Page 42 Buyer or other terms and conditions relating to the use of the Purchased Device or Licensed Process provided to Buyer by Lonza, or (c) the gross negligence or willful mis- conduct of Buyer or any of its affiliates in performing any activity relating to the subject matter hereof.
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Page 43: Technical Specifications
96-well Unit: 4.7 kg (10.4 lb) 24.5 x 10.5 x 28 (cm)/ 9.7 x 4.1 x 11 (in)/ 9.7 x 8.1x 11 Hersteller / supplier / manufacture Lonza Cologne GmbH Nattermannallee 1 D-50829 Cologne, Germany Herstellungsdatum / date of manufacture /... - Page 44 Nucleocuvette, maxGFP and 96-well Shuttle are trademarks of Lonza Cologne GmbH in Germany and / or the U.S. and / or other countries.All trademarks belong to Lonza, registered in USA, EU or CH or to third party own- ers and used only for informational purposes. The information contained herein is believed to be correct and corresponds to the latest state of scientific and technical knowledge.
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