Agilent Seahorse XF User Manual
Seahorse XF HS Mini Extracellular Flux Analyzer
User Manual
Notices
CAUTION
WARNING
© Agilent Technologies, Inc. 2020
No part of this manual may be reproduced in
any form or by any means (including electronic
storage and retrieval or translation into a foreign language) without prior agreement and
written consent from Agilent Technologies,
Inc. as governed by United States and international copyright laws.
Manual Part Number
5994-1961EN
Edition
First edition, May 2020
Printed in USA
Agilent Technologies, Inc.
121 Hartwell Ave.
Lexington, MA 02466
Manufactured by:
Agilent Technologies, Inc.
300 Griffith Rd,
Chicopee, MA 01022
Warranty
The material contained in this document is
provided “as is,” and is subject to being
changed, without notice, in future editions.
Further, to the maximum extent permitted by
applicable law, Agilent disclaims all warranties, either express or implied, with regard to
this manual and any information contained
herein, including but not limited to the implied
warranties of merchantability and fitness for
a particular purpose. Agilent shall not be liable for errors or for incidental or consequential damages in connection with the
furnishing, use, or performance of this document or of any information contained herein.
Should Agilent and the user have a separate
written agreement with warranty terms covering the material in this document that conflict with these terms, the warranty terms in
the separate agreement shall control.
Technology Licenses
The hardware and/or software described in this
document are furnished under a license and
may be used or copied only in accordance with
the terms of such license.
Restricted Rights Legend
U.S. Government Restricted Rights. Software
and technical data rights granted to the federal
government include only those rights customarily provided to end user customers. Agilent
provides this customary commercial license in
Software and technical data pursuant to FAR
12.211 (Technical Data) and 12.212 (Computer
Software) and, for the Department of Defense,
DFARS 252.227-7015 (Technical Data -Commercial Items) and DFARS 227.7202-3 (Rights
in Commercial Computer Software or Computer Software Documentation).
Safety Notices
A CAUTION notice denotes a
hazard. It calls attention to an
operating procedure, practice, or the
like that, if not correctly performed
or adhered to, could result in
damage to the product or loss of
important data. Do not proceed
beyond a CAUTION notice until the
indicated conditions are fully
understood and met.
A WARNING notice denotes a
hazard. It calls attention to an
operating procedure, practice, or the
like that, if not correctly performed
or adhered to, could result in
personal injury or death. Do not
proceed beyond a WARNING notice
until the indicated conditions are
fully understood and met.
Contents
1Introduction
Safety Considerations 5
Overview 6
Technical Specifications 7
2 Installation
Unpacking and Component Identification 9
Suitable Locations for the XF HS Mini Analyzer 10
3 Basic Operations
Power and Warm Up 11
Power up 11
Welcome screen on first time power up 11
XF HS Mini Analyzer Communications and Network Connection Setup 12
Turning the XF HS Mini Analyzer Off 12
4 Navigating the XF HS Mini Analyzer
Setting Up an XF HS Mini Assay 14
Running an XF HS Mini Assay 18
Modifying XF HS Mini Analyzer Settings 22
XF HS Mini Analyzer system files 23
System files 23
Diagnostics tests 24
System settings 26
Assay results 31
Template management 33
Help 33
Using Safe Lock (Trend Micro) 35
Cleaning and Routine Maintenance 36
Air filter replacement 36
Analyzing XF HS Mini Analyzer Data Using Seahorse Analytics 37
5 XF HS Mini Analyzer Network Setup
Introduction 42
Why should I network my XF HS Mini Analyzer? 42
Network access features on the XF HS Mini Analyzer 43
XF HS Mini Analyzer System Information 44
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 3
Required materials 44
Wired Network Setup 45
Wired connection 45
Wireless Network Setup 46
Wireless connection 46
Joining a wireless network 47
Shared folder setup 48
Email configuration 49
Network Checklist 52
6Support
Troubleshooting Guide 53
Technical Support and Ordering Information 54
Worldwide technical support 54
Ordering 54
Online help and support 55
Additional Resources 56
4 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
1 Introduction
WARNING
Safety Considerations 5
Overview 6
Technical Specifications 7
Safety Considerations
WARNING
The protection provided by this instrument may be compromised if it is used in a manner not
specified by Agilent.
Safe operation of the Agilent Seahorse XF HS Mini Analyzer requires all covers to be securely
attached and the plate tray door to be closed. This also prevents heat loss and system cooling,
which can affect data quality.
The door opens automatically when the tray is extended, allowing the operator to insert or remove
the assay consumables. Exercise caution when loading the miniplate/cartridge to avoid the
possibility of a pinch hazard. After the miniplate/cartridge is placed securely on the tray, the
remove your hand from the tray area before continuing the assay. After the command is given to
continue the assay through the user interface, the tray will move slowly back into the instrument,
and the door will close. An optical sensor is used to determine the status of the door.
The XF HS Mini Analyzer has heaters around the miniplate that maintain a stable tray
temperature. The tray temperature is maintained at 37 °C, and monitored by temperature sensors
and controllers embedded above the tray. A thermal fuse disables the heater when it reaches an
abnormally high tray temperature.
Do not replace the power cord provided with any other power cord that is rated at less than what
is specified in “Technical Specifications” on page 7. This depends on the power mains of the
country where the instrument will be used.
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 5
Introduction
Overview
The XF HS Mini Analyzer measures the rate of change in dissolved oxygen and pH in the media
immediately surrounding living cells cultured in a miniplate. Changes in the extracellular media are
caused by cell consumption or production of analytes. A sensitive measurement of the media flux
can be used to determine rates of cellular metabolism with precision and in a noninvasive,
label-free manner.
A unique feature of XF technology is its ability to make accurate and repeatable measurements in
as little as five minutes. This is accomplished by isolating an extremely small volume
(approximately 2 µL) of media above the cell monolayer. Cellular metabolism causes rapid, easily
measured changes to the microenvironment in this small volume.
A measurement cycle is performed in six minutes. The media is gently mixed and the analyte
levels are then measured until the oxygen concentration drops approximately 20 to 30%, and
media pH declines approximately 0.1 to 0.2 pH units. The measurement is performed using
optical fluorescent biosensors embedded in a disposable cartridge. The cartridge is placed into
the XF HS Mini Analyzer cell culture miniplate.
Baseline metabolic rates are typically measured three to four times, and are reported in pmol/min
for oxygen consumption rate (OCR) and in mpH/min for extracellular acidification rate (ECAR).
The compound is then added to the media and mixed, and then the post-treatment OCR and
ECAR measurements are made and repeated. As cells shift metabolic pathways, the relationship
between OCR and ECAR changes.
The XF HS Mini Analyzer system includes:
• Benchtop analyzer
• Disposable sensor/compound
• Delivery cartridges
• Tissue-culture treated miniplates for seeding cells for analysis
• Calibration solution
• Seahorse Analytics for XF data analysis
Consumables are sold separately and include FluxPaks (sensor cartridges, cell plates, and
calibrant) as well as various assay kits, reagents, and media. Seahorse XF HS Mini sensor
cartridges are available exclusively from Agilent.
6 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
Introduction
S7852A
Technical Specifications
Model Seahorse XF HS Mini Analyzer (model number S7852A)
Dimensions Width × height × depth
12 in × 17 in × 23 in
30 cm × 43 cm × 58 cm
Weight 33 lb/15 kg
Power requirements 100-240 VAC 50/60 Hz
6 A/3 A
Power cord rating 3-wire (grounded) AC power cord rated 10 A or greater
Power fuse ratings 250 V/5 A time lag (two fuses) 5 mm × 20 mm
Environmental operating range +39 °F - 86 °F/+4 °C - 30 °C
No direct sunlight
Humidity 20 to 70% RH, noncondensing
Sample temperature and
environment
Data interface TCP/IP (external)
Software OS Windows 10 embedded
Controlled to user-selected temperature between 16 and 40 °C, but at least 10 °C above
ambient temperature
No gas or humidity control
USB type A (one in front, two in back)
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 7
Introduction
8 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
2 Installation
Unpacking and Component Identification 9
Suitable Locations for the XF HS Mini Analyzer 10
This chapter provides unpacking and installation instructions for the Agilent Seahorse XF HS Mini
Analyzer.
Unpacking and Component Identification
After receiving the package, immediately check each box for damage. Report any shipping
damage to the transportation company, and Agilent using “Technical Support and Ordering
Information” on page 54. Table 1 shows the Seahorse XF HS Mini Analyzer system components.
Table 1 Seahorse XF HS Mini Analyzer system components
Instrument/Controller Quantity Image
XF HS Mini Analyzer 1
Power cord (region specific) 1
Wireless Micro USB adapter
(p/n S7802-80000)
1
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 9
Installation
Suitable Locations for the XF HS Mini Analyzer
The XF HS Mini Analyzer is designed for laboratory use. The internal environment for the
XF HS Mini Analyzer cell culture miniplate is controlled to a preset tray temperature. The
laboratory room temperature must therefore be maintained within the range listed in “Technical
Specifications” on page 7. The miniplate tray temperature can be monitored using the status
display in the upper right portion of the user interface.
The XF HS Mini Analyzer uses optical detection technology to measure extremely low levels of
fluorescent emission from analyte sensors. Although the instrument has been designed to shield
room light, avoid excessive light (such as direct sunlight).
10 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
3 Basic Operations
Power and Warm Up 11
XF HS Mini Analyzer Communications and Network Connection Setup 12
Turning the XF HS Mini Analyzer Off 12
This chapter provides basic operating procedures for the Agilent Seahorse XF HS Mini Analyzer.
Power and Warm Up
Power up
To power on the XF HS Mini Analyzer:
1 Press the power switch on the back of the instrument. (Figure 1)
Figure 1. XF HS Mini Analyzer power switch.
2 Allow at least 20 minutes for the instrument to fully warm and equilibrate to the set
temperature.
3 Check the status icon in the upper right corner of the screen. When ready the tray temperature
and a green check mark is displayed.
Welcome screen on first time power up
During the first power up, a welcome screen displays initial diagnostic tests on the instrument.
Follow the on-screen directions to run them.
Run these tests at any time using the Diagnostics menu selection on the instrument user
interface. For more details, see “Navigating the XF HS Mini Analyzer” on page 13.
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 11
Basic Operations
NOTE
NOTE
XF HS Mini Analyzer Communications and Network Connection Setup
The USB connectors on the instrument transfer template and result files between the XF HS Mini
Analyzer and a computer running Seahorse Analytics. Agilent recommends connecting the XF HS
Mini Analyzer to a local network directory to ease file movement and aid Cell Analysis Technical
Support in servicing the instrument. Please contact Cell Analysis Technical Support, “Worldwide
technical support” on page 54, with any questions.
For instructions on setting up a wired network connection, see “Wired Network Setup” on
page 45. For instructions on setting up a wireless network connection using the included wireless
adapter, see “Wireless Network Setup” on page 46.
NOTE
Only the included wireless adapter has been qualified to work with the XF HS Mini Analyzer.
?
Turning the XF HS Mini Analyzer Off
To turn off the instrument, press the power button in the lower-left corner of the Home screen to
first shut down the XF HS Mini Analyzer screen. After the screen turns black, use the power switch
on the back of the instrument to completely shut down power.
NOTE
Shut down the software before powering off using the rear power switch.
12 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
4 Navigating the XF HS Mini Analyzer
Setting Up an XF HS Mini Assay 14
Running an XF HS Mini Assay 18
Modifying XF HS Mini Analyzer Settings 22
XF HS Mini Analyzer system files 23
Help 33
Using Safe Lock (Trend Micro) 35
Cleaning and Routine Maintenance 36
Analyzing XF HS Mini Analyzer Data Using Seahorse Analytics 37
This chapter provides navigation information for the Agilent Seahorse XF HS Mini Analyzer.
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 13
Navigating the XF HS Mini Analyzer
Setting Up an XF HS Mini Assay
Select a template from local, USB, or network drive
Agilent provides default templates for most Seahorse XF assay kits. Templates can also be
created in Wave desktop on a desktop or laptop computer and then transferred to the XF HS Mini
Analyzer through a network connection or USB flash drive.
1 Click Start from the Home screen. See Figure 2.
Figure 2. Home screen.
2 Select a template provided by Agilent to perform a specific assay. These templates can be
found in the Local tab. See Figure 3.
Figure 3. Local tab.
14 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
Navigating the XF HS Mini Analyzer
NOTE
3 Templates designed in Wave can be transferred to the XF HS Mini Analyzer through a USB or
network location.
• If the instrument is networked, click the Network tab, select and run the template from the
networked location.
• If a USB drive containing a valid XF HS Mini Analyzer template is inserted into the
instrument, click the USB tab, select and run the template directly from the USB drive.
NOTE
Only XF HS Mini Analyzer assay template files (*.asyt) are recognized by the XF HS Mini
Analyzer. Templates must reside on the root of the USB flash drive and not in a subfolder.
Verify groups and plate map
After selecting the template file, make any necessary modifications to the plate map and groups
to be analyzed.
1 To see the conditions defined for a group, click the group name and look at the header
information, as shown in Figure 4.
Figure 4. Groups.
2 To change the wells assigned to each group, click the group name followed by the well or
wells to be included.
3 For all other changes to Groups, use Seahorse Analytics.
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 15
Navigating the XF HS Mini Analyzer
Review/edit instrument protocol
1 Check that all desired steps are selected, check boxes checked (Figure 5). Agilent strongly
recommends performing equilibration for all cell-based XF assays.
2 If needed, increase or decrease the number of measurements that will be performed during
the assay on the Instrument Protocol page.
a Click the number in the cycle column that corresponds with the step to modify.
b Use the arrows to adjust the number of cycles up or down. See Figure 5.
Figure 5. Instrument protocol page.
Review summary and start assay
1 Review the “Safety Considerations” on page 5 to verify the desired settings.
2 Before beginning an assay, the following optional steps may be taken:
a Click the Assay Result File Name text box to customize the name of the assay result file.
See Figure 6.
Figure 6. Assay name.
b Click Project Notes to add any notes related to the assay or protocol being performed.
16 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
Navigating the XF HS Mini Analyzer
NOTE
c Click Email Notification to add email addresses for recipients to be notified when the
assay is complete (requires an active network connection).
3 Click Start Run then follow the onscreen instructions.
NOTE
The XF HS Mini Analyzer will save assay results to a USB flash drive by default (if available).
Agilent recommends inserting a USB flash drive into the instrument at the beginning of assay
setup so that results will automatically be saved. If the XF HS Mini Analyzer does not detect a
USB flash drive after completing an assay, the system will prompt the user to insert a USB to
save their assay result file.
Seahorse XF HS Mini Extracellular Flux Analyzer User Manual 17
Navigating the XF HS Mini Analyzer
NOTE
Running an XF HS Mini Assay
1 Load the cartridge (hydrated and loaded with compounds) and utility plate onto the tray when
prompted.
2 Ensure the cartridge fits properly on the utility plate, the lid is removed from the cartridge, and
the direction of the cartridge matches the image on the screen. See Figure 7.
Figure 7. Load cartridge screen.
3 The XF HS Mini Analyzer will perform calibration of the sensor cartridge. This takes
approximately 20 minutes.
NOTE
Although the sensor calibration may appear complete after 15 minutes, the instrument will be
busy for a few more minutes. Wait for the user prompt to appear.
4 The XF HS Mini Analyzer tray will open and present the utility plate. Remove the utility plate
and load the cell plate. Ensure the lid is removed from the cell plate before loading onto the XF
HS Mini Analyzer tray.
5 Click Continue to begin the equilibration step. The progress bar in the upper-left area of the
screen displays the progress of the equilibration step.
6 Click either the Overview or OCR vs. ECAR tab to toggle between the views during the assay.
See Figure 8 on page 19. Overview and OCR versus ECAR are the two run-time views available
as data is acquired by the XF HS Mini Analyzer.
18 Seahorse XF HS Mini Extracellular Flux Analyzer User Manual
Loading...