ThermoFisher Scientific Invitrogen NEON1 User manual

For Research Use Only. Not for use in diagnostic procedures.
Neon™NxT Electroporation System
USER GUIDE
For electroporation of mammalian cells, including primary and
stem cells, with high transfection eciency
Catalog NumbersNEON1, NEON1SK, NEON1P, NEON1PS
Publication Number MAN0026677
Revision A.0

Life Technologies Holdings Pte Ltd | Block 33 | Marsiling Industrial Estate Road 3 | #07-06, Singapore 739256
For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition.
Revision history:Pub.No. MAN0026677
Revision Date Description
A.0 22 March 2023 New product manual for the Neon™NxT Electroporation System.
The information in this guide is subject to change without notice.
DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE
LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR
ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these
products, you accept the terms and conditions of all applicable Limited Use Label Licenses.
TRADEMARKS: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.
©2023 Thermo Fisher Scientific Inc. All rights reserved.

Contents
■CHAPTER1Productinformation .................................................. 6
Productdescription ............................................................. 6
System overview ........................................................... 6
Features ................................................................... 7
Product contents ................................................................ 7
Neon™NxT System contents ................................................. 7
Neon™NxT Kit contents ..................................................... 8
Systemcomponents ............................................................. 9
Neon™NxTdevice .......................................................... 9
Neon™NxT Pipette Station .................................................. 10
Neon™NxT Pipette ........................................................ 11
Neon™NxTKit ............................................................ 11
Neon™NxT Tip ............................................................ 11
Neon™NxT Tube .......................................................... 12
Upon receiving thedevice ....................................................... 12
Unpackinginstructions ..................................................... 12
Getting started ................................................................ 13
Set up the Neon™NxT Electroporation System ................................ 13
User interface overview ..................................................... 16
Touchscreen controls ...................................................... 16
Connect the instrument to the Internet ....................................... 19
Create a user profile on the instrument ....................................... 23
Manage userprofiles ....................................................... 23
About the Thermo Fisher™ConnectPlatform ...................................... 25
Features .................................................................. 25
Create a Connect Platformaccount .......................................... 25
Create a PINnumber ....................................................... 25
Set up a new administrator .................................................. 26
Add an instrument to your Connect Platformaccount .......................... 26
Neon™NxT Electroporation System User Guide 3

■CHAPTER2Methods ............................................................. 28
Electroporation protocoloptions ................................................. 28
Electroporation parameters ................................................. 28
Create or edit protocols ......................................................... 29
Create a protocol .......................................................... 29
Edit a protocol ............................................................ 30
Save a Protocol ........................................................... 30
Create or edit platemaps ....................................................... 31
Create a platemap ......................................................... 31
Edit a platemap ........................................................... 32
Save a platemap .......................................................... 33
Manage protocols .............................................................. 33
Import or export protocols .................................................. 33
Generalguidelines ............................................................. 37
Recommendedkits ........................................................ 37
Recommendedbuers ..................................................... 38
DNA quality andamount .................................................... 38
siRNA quality andamount .................................................. 38
Controls .................................................................. 38
Using the Neon™NxT Electroporation System ..................................... 39
Required materials notsupplied ............................................. 39
Calculate electroporation volumes for cell suspension ......................... 40
Prepare adherentcells ...................................................... 40
Prepare suspensioncells ................................................... 42
Set up the pipettestation ................................................... 43
Load the pipette ........................................................... 43
Run electroporation protocol ................................................ 45
Maintenance .................................................................. 47
Cleaning and maintenance .................................................. 47
Replace thefuses .......................................................... 48
Upgrade the systemfirmware ............................................... 48
Self Verification test ........................................................ 50
Restore factory settings (Administrator profile only)............................ 51
Repackaging theinstrument ................................................ 51
■APPENDIXATroubleshooting .................................................... 53
Instrument errorcodes .......................................................... 55
■APPENDIXBOptimization ........................................................ 60
Optimization protocol ........................................................... 60
Contents
4Neon™NxT Electroporation System User Guide

■APPENDIXCRecommended instrumentsettings .............................. 61
AboutInstrument .............................................................. 61
Recommended instrumentsettings .............................................. 61
Instrument settings (Administrator only) ...................................... 62
Maintenance & services ......................................................... 65
Run History ................................................................... 66
■APPENDIXDSpecifications ...................................................... 68
Productspecifications .......................................................... 68
Networking requirements ....................................................... 69
■APPENDIXERelated products ................................................... 70
Accessory products ............................................................ 70
Additional products ........................................................ 70
■APPENDIXFSafety ............................................................... 78
Safetyinformation .............................................................. 78
Informationalsymbols .......................................................... 78
Informations de sécurité ........................................................ 80
Informationalsymbols .......................................................... 80
Environmental requirements ..................................................... 82
Safety and electromagnetic compatibility (EMC) standards .......................... 83
Safety .................................................................... 83
EMC ..................................................................... 83
Class Anotice ............................................................. 83
Environmental design standards ............................................. 84
Electricalsafety ................................................................ 84
Service operation requirements .............................................. 84
Chemicalsafety ................................................................ 86
Biological hazardsafety ......................................................... 87
■APPENDIXGDocumentation and support ...................................... 88
Customer and technical support ................................................. 88
Limited product warranty ........................................................ 88
Contents
Neon™NxT Electroporation System User Guide 5

Product information
Product description
The Neon™NxT Electroporation System is a benchtop electroporation device that employs an
electroporation technology which uses a pipette tip as an electroporation chamber to eciently
transfect mammalian cells including primary and immortalized hematopoietic cells, stem cells, and
primary cells.
The Neon™NxT Electroporation System eciently delivers nucleic acids, proteins, and siRNA into all
mammalian cell types with a high cell survival rate. The transfection is performed using as few as 1 ×
104or as many as 5 × 106cells per reaction in a volume of 10 µL or 100 µL for a variety of cell culture
formats (60mm, 6-well, 48-well, 24-well, and 96-well).
The Neon™NxT Electroporation System uses a single transfection kit (Neon™NxT Kit) that is compatible
with various mammalian cell types including primary and stem cells thereby avoiding the need to
determine an optimal buer for each cell type.
The Neon™NxT Electroporation System uses open and transparent protocols that are optimized for
ease of use and simplicity. The Neon™NxT device is preprogrammed with 24 optimization protocols,
and over 300 preprogrammed cell-specific protocols. In addition, there is capacity for 10,000 more
protocols in the Neon™NxT device database. Optimized protocols for many commonly used cell types
are also available at thermofisher.com/transfectionprotocolsandcitations to maximize transfection
eciencies for your cell types.
See “System components” on page9 for details on various parts of the system.
System overview
Unlike standard cuvette based electroporation, the Neon™NxT Electroporation System uses a unique
electroporation reaction chamber, the Neon™NxT Tip that delivers a high electric field to the biological
sample. The Neon™NxT Tip maximizes the gap size between the two electrodes while minimizing
the surface area of each electrode. As a result, the sample experiences a more uniform electric field,
minimal pH change, less ion formation, and negligible heat generation.
The established Neon™electroporation technology overcomes limitations associated with standard
cuvette based electroporation to increase transfection eciency and cell viability, and provide an
ergonomic workflow.
1
6Neon™NxT Electroporation System User Guide

Features
Important features of the Neon™NxT Electroporation System are listed below:
•Small footprint—the benchtop design fits inside a tissue culture hood, reducing contamination risk.
•Preserves samples—minimize sample transfer loss with the complete elimination of an
electroporation cuvette and associated pipetting steps.
•Flexible—deliver DNA, RNA, and protein to 1 × 104to 5 × 106cells per reaction in a sample volume
of 10µL or 100µL.
•Customizable—optimize electroporation parameters freely, or leverage over 300 preprogrammed
protocols with the ability to save up to 10,000 more protocols.
•Simplicity—easy to perform workflow with only three steps; a single reagent kit is all that is needed
for most cell types and delivery payloads.
•Connectivity—the TransfectionLab app paired with the Thermo Fisher™Connect Platform
(https://apps.thermofisher.com/apps/spa/#/dashboard) allows for digital experiment design and
connectivity to the Neon™NxT device.
•Improved usability—pipette ergonomics with ClipTip technology for loading and unloading of the
pipette tips, enhanced feedback loop, and intuitive user interface with plate setup.
Product contents
Neon™NxT System contents
The contents of the Neon™NxT Electroporation System are listed in the following table. The Neon™NxT
Electroporation System is shipped at room temperature.
See page9 for descriptions of the Neon™NxT Electroporation System, and page28 to set up the
device.
Product Quantity
Neon™NxT Electroporation Device 1
Power Cord
(for US/Canada/Taiwan/Japan, Europe, or UK) 1
Neon™NxT Pipette 1
Neon™NxT Pipette Station 1
Quick Reference 1
Chapter1Product information
Product contents 1
Neon™NxT Electroporation System User Guide 7

Neon™NxT Kit contents
Neon™NxT Kits are used with the Neon™NxT Electroporation System for ecient transfection of
mammalian cells and are available as standalone products (see “Accessory products” on page70).
The kits consist of two components which are not sold individually (a Tips/Tubes Kit, and a Buer Kit),
and are available in two formats (for electroporation of 10µL samples, and 100µL samples).
Neon™NxT Kit components are listed in the following table, and are shipped at room temperature.
After receiving the kit, store buers at 4℃and tips/tubes at room temperature.
Catalog numbers that appear as links open the web pages for those products.
Item
Neon™NxT Kit, 10µL Neon™NxT Kit, 100µL
Cat. No. N1025
(50 reactions)
Cat. No. N1096
(192 reactions)
Cat. No. N10025
(50 reactions)
Cat. No. N10096
(192 reactions)
Tips/Tubes Kit N1025K N1096K N10025K N10096K
Neon™NxT Tips 25 tips (10µL) 96 tips (10µL) 25 tips (100µL) 96 tips (100µL)
Neon™NxT Tubes 8 32 8 32
Buer Kit N1025R N1096R N10025R N10096R
Neon™NxT Resuspension
R Buer (Proprietary) 1mL 4 × 1mL 10mL 4 × 10mL
Neon™NxT Resuspension T
Buer (Proprietary) 1mL 4 × 1mL 10mL 4 × 10mL
Neon™NxT Electrolytic E10
Buer (Proprietary) 50mL 2 × 100mL — —
Neon™NxT Electrolytic
E100 Buer (Proprietary) — — 50mL 2 × 100mL
Chapter1Product information
Product contents
1
8Neon™NxT Electroporation System User Guide

System components
Neon™NxT device
The electrical parameters of the Neon™NxT device are factory set and do not require calibration. It is
used with the Neon™NxT Pipette Station and Neon™NxT Kits to eciently transfect mammalian cells
including primary and stem cells.
Front and rear view
2
1
1Status indicator LED
2Touchscreen
1
2
43
98
5
5
7
6
1Power switch
2AC inlet (connect to the power cord, and plug into a
power outlet)
3High voltage port (connect to the high voltage
connector on one end of the power cord)
4Ethernet port (covered port; use a hex key for access)
5USB 2.0 port (covered port; use a hex key for access)
6Hex key (Allen wrench)
7Connector cable storage
8USB wireless adaptor port
9Low voltage interface port (connects to the low voltage
interface connector on one end of the power cord)
Chapter1Product information
System components 1
Neon™NxT Electroporation System User Guide 9

LED status indicators
LED color Status
Blue Instrument is idle.
Green Instrument is in operation.
Blinking green Instrument high voltage subsystem in operation.
Blinking amber Warning/error
About the covered ports
Do not access the covered ports at the rear of the device during electroporation. The ports should
remain covered during the protocol, and should only be accessed when the device is in idle mode.
To open the covers over the ports, place one end of the supplied wrench into the hole located in the
cover and slide it down to expose the port.
Neon™NxT Pipette Station
The Neon™NxT Pipette Station holds a Neon™NxT Pipette during electroporation procedures. The
pipette (with attached tip) is held in the Neon™NxT Tube which has an electrode near the bottom that
transfers the electric field from the electrode inside the Neon™NxT Tip.
5
6
7
1 3
4
2
2
1
1Neon™NxT Pipette Station
2Tube chamber for Neon™NxT Tube
3Low voltage interface port
4High voltage interface port
5Connector cable
6Low voltage interface connector
7High voltage interface connector
Chapter1Product information
System components
1
10 Neon™NxT Electroporation System User Guide

Neon™NxT Pipette
The Neon™NxT Pipette utilizes a positive displacement
pipette mechanism for pipetting mixtures containing cells
and payload (such as nucleic acid). The Neon™NxT Pipette
is a fixed volume pipette and permanently calibrated at
the manufacturing stage and does not require any further
calibration.
The Neon™NxT Pipette is designed for use with Neon™
NxT Tips only. Do not use any other tips with the Neon™
NxT Pipette.
Neon™NxT Kit
Neon™NxT Kits contain Neon™NxT Tips, Neon™NxT Tubes, and buers for electroporation. Neon™
NxT Kits are available in two formats for electroporation of 10µL or 100µL samples, as well as in two
reactions sizes of 25 × 2 reactions and 96 × 2 reactions (See page 70 for ordering information).
Neon™NxT Tip
Neon™NxT Tips are disposable tips composed of a tip
and piston used with the Neon™NxT Pipette. Neon™NxT
Tips contain a gold-plated electrode to create a disposable
electric chamber for the delivery of a high electric field
to biological samples. The tips are supplied with Neon™
NxT Kits in two formats to support operating volumes of
10 µL and 100 µL, respectively (see page 70 for ordering
information).
To ensure repeatability and eliminate variation of the
transfection conditions within or between experiments,
do not use the tip more than 2times. Oxide formation
at the piston surface area can be generated if the tips
are used more than 2 times, which decreases electrode
function of the piston. Change the tip when switching to a
dierent payload or cell type.
Tip specifications:
Material (tip housing): Polypropylene
Capacity: 10µL or 100µL
1
2
3
1Plunger
2Tip ejector
3Tip holder
2
1
10-μL 100-μL
1Gold plated plunger/piston (raised out of tip
housing)
2Tip housing
Chapter1Product information
System components 1
Neon™NxT Electroporation System User Guide 11

Neon™NxT Tube
The Neon™NxT Tube holds electrolytic buer during
electroporation and is inserted into the Neon™NxT Pipette
Station. The Neon™NxT Pipette with the Neon™NxT Tip
is then inserted into the Neon™NxT Tube which has an
electrode near the bottom that transfers the electric field
from the electrode inside the Neon™NxT Tip. Neon™NxT
Tubes are supplied with Neon™NxT Kits as well as available
separately (see page 70).
To avoid contamination, do not use tubes more than
12times. Change the tube and buer when switching to a
dierent payload or cell type.
Tube specifications:
Material: Polycarbonate
Capacity: 2–3mL
Upon receiving the device
Examine the unit carefully for any damage incurred during transit. Any damage claims must be
filed with the carrier. If the instrument is damaged during shipment, contact Technical Support for
assistance. To register the device, activate your warranty, and be notified of important updates, go to
thermofisher.com.
Unpacking instructions
Consult the following instructions to unpack the Neon™NxT Electroporation System. The weight of the
system is 13pounds (5.9kg).
1. Open the shipping box to unpack the Neon™NxT device. Save the box and other packaging
material (in case you need to transport or ship the unit).
2. Remove the packing material, then inspect the Neon™NxT device and Neon™NxT Pipette Station
for shipping damage.
3. Remove the Neon™NxT device and the Neon™NxT Pipette Station from the box and place them
on a flat, level surface.
4. Set up the Neon™NxT Electroporation System as described on page 13.
2
4
3
5
1
1Electrode
2Buer
3Buer level indicator line
4Hook profile
5Raised dots to assist grip
Chapter1Product information
Upon receiving the device
1
12 Neon™NxT Electroporation System User Guide

Getting started
Set up the Neon™NxT Electroporation System
1. Unpack the Neon™NxT Electroporation System as described in “Unpacking instructions” on
page12.
2. Place the Neon™NxT device on a level laboratory bench. Keep an area of free space at least 10
cm surrounding the unit to ensure proper ventilation.
Note: The Neon™NxT device has a small footprint and can be easily set up in the tissue culture
hood for convenience.
IMPORTANT! Position the device properly such that the power switch and AC inlet located at the
rear of the unit (see “Front and rear view” on page9) are easily accessible. Be sure to position the
device such that it is easy to disconnect the unit.
Since Neon™NxT device is air-cooled, its surface may become hot during operation. When
installing the device, leave a space of more than 10 cm from the back of the device.
3. Place the Neon™NxT Pipette Station near the Neon™NxT device.
4. Connect the high voltage and low voltage interface connectors on the cable to the high voltage
and low voltage interface ports at the rear of the Neon™NxT Pipette Station.
IMPORTANT! Always handle the connectors using the cord plug and not the cord cable when
connecting or disconnecting the connectors.
Chapter1Product information
Getting started 1
Neon™NxT Electroporation System User Guide 13

5. Connect the high voltage and low voltage interface connectors on the Neon™NxT Pipette Station
to high voltage and low voltage interface ports at the rear of the Neon™NxT device.
Be sure to align the ridge on the connector of the cable with the groove on the port of the Neon™
NxT device.
IMPORTANT! Always handle the connectors using the cord plug and not the cord cable when
connecting or disconnecting the connectors.
6. Ensure the AC power switch is in the O position ( see “Front and rear view” on page9).
Chapter1Product information
Getting started
1
14 Neon™NxT Electroporation System User Guide

7. Attach the power cord to the AC inlet at the rear of the Neon™NxT device, then plug the cord into
an electrical outlet. Use only properly grounded AC outlets and power cords.
The instrument operates at voltages of 100–240 VAC and the frequency range of 50/60 Hz. Ensure
that the local supply voltage in the laboratory conforms to that specified on the type label on the
back of the instrument
8. Press the main power switch at the rear of the device to the ON position to turn on the power.
9. The Neon™NxT device is operated by the digital touch screen at the front of the device. See “User
interface overview” on page16 for details.
After the Neon™NxT Electroporation System is set up, see “Using the Neon™NxT Electroporation
System” on page39 for instructions on performing electroporation.
Chapter1Product information
Getting started 1
Neon™NxT Electroporation System User Guide 15

User interface overview
Symbol Function
Main dial
•Set up run
•Select a protocol or plate map
•Create a new protocol (see “Create or edit protocols” on page29 for more details)
•Create a new plate map (see “Create or edit plate maps” on page31 for more details)
Displays the instrument status when a protocol is run (see “Touchscreen status indicators”
on page17)
Optimization screen
•View protocols
•Create a new protocol (see “Create or edit protocols” on page29 for more details)
•Edit an existing protocol (see “Create or edit protocols” on page29 for more details)
Create protocol screen
•Calculate volume for protocol (see “Calculate electroporation volumes for cell
suspension” on page40 for more details)
Touchscreen controls
Table1General touchscreen controls
Button Function
Returns to the previous screen.
Go to Home screen.
Go to Sign in screen.
Go to Settings screen.
Close the current modal window.
Chapter1Product information
Getting started
1
16 Neon™NxT Electroporation System User Guide

Touchscreen status indicators
Table2Status indicators
Button Function
Indicates whether a USB device is inserted into the
instrument.
Indicates whether the Wi-Fi is on or o.
Indicates whether the instrument is connected to wired
network.
Indicates whether the instrument is connected to the
Thermo Fisher™Connect Platform.
Enter text
When you press a field that requires the input of text, the text editor, as seen in the following figure,
opens.
23 5 4
16
1Enter a letter
2Change letter case
3Enter punctuation or other symbols
4Delete
5Close and save
6Close without saving
Chapter1Product information
Getting started 1
Neon™NxT Electroporation System User Guide 17

Enter numbers
When you press a field that requires a numerical input, the numeric editor, as seen in the following
figure, opens.
23
4
1
1Enter a number
2Delete/backspace
3Close and save
4Close without saving
Chapter1Product information
Getting started
1
18 Neon™NxT Electroporation System User Guide

Connect the instrument to the Internet
The instrument can connect to the Internet by either wired or wireless methods.
To connect by wired method through the instrument Ethernet port using a cable, see “Set up a wired
connection” on page19.
To connect by wireless method through the instrument USB wireless adapter port with a USB-enabled
Wi-Fi card, see “Set up a wireless connection” on page22.
Set up a wired connection
Connect one end of a Ethernet cable to the instrument Ethernet port, and the other end to an Ethernet
port wall plug (see “Front and rear view” on page9 for port location).
1. On the Home screen, select (Settings).
2. In the Settings screen, select Instrument Settings.
Chapter1Product information
Getting started 1
Neon™NxT Electroporation System User Guide 19

3. In the Instrument Settings screen, select Network configuration.
4. In the Network Connection screen, select a field in the Wired panel.
1 2
1Wireless panel 2Wired panel
5. Select a method to enter an IP address.
a. Select DHCP to obtain an IP address automatically. A check mark appears when DHCP is
selected.
Chapter1Product information
Getting started
1
20 Neon™NxT Electroporation System User Guide