Agilent Technologies Seahorse XFe96 - User Manual

Basic Procedure
A key feature of the Agilent Seahorse XFe96/XF96 Analyzer is the ability to inject
solutions during the assay and observe changes in respiration or glycolytic rates in
real time. This is accomplished by pipetting solutions into the Injection Ports on the
XF Sensor Cartridge prior to beginning an XF assay. This procedure describes the
injection port loading process which is performed the day of your assay following
overnight XF Sensor Cartridge hydration.
Recommended injection volume is 20-30 µL for XFe96/XF96 Analyzers.
Recommended Injection Solution Volumes for 10X dilution upon injection, starting
with a microplate well volume of 180 µL assay medium (Figure 1):
1. Port A: 20 µl
2. Port B: 22 µl
3. Port C: 25 µl
4. Port D: 28 µl
The composition and number of ports utilized will depend on your assay design.
Loading the Agilent Seahorse XFe96
Sensor Cartridge Injection Ports
User Guide
Seahorse
A
C
E
G
B
D
F
H
A B
C D
Port designation
for each well
Figure 1. Recommended injection volumes and port designation for each well.

Summary

Loading the Agilent Seahorse XFe96 Sensor Cartridge Injection Ports

Basic Procedure

Describes the injection port loading process, following overnight XF Sensor Cartridge hydration.

Requirements for Injection Port Loading

Loading the XFe96 Sensor Cartridge with Injection Solutions

Step 1: Prepare and Warm Injection Solutions

Prepare and warm injection solutions to 37°C, ensuring correct pH for accurate results.

Step 2: Orient the XFe96 Sensor Cartridge

Properly orient the XFe96 Sensor Cartridge on the work surface for loading.

Step 3: Place the A;D Port Loading Guide

Position the A/D port loading guide correctly on the sensor cartridge for stable loading.

Step 4: Dispense Injection Solutions

Dispense injection solutions using a multi-channel pipette with correct technique.

Step 5: Withdraw Tips and Remove Loading Guide

Carefully withdraw pipette tips and remove the loading guide after dispensing.

Step 6: Load B;C and D Ports

Switch and use the B/C port loading guide for subsequent port loading.

Step 7: Visual Inspection and Analyzer Transfer

Perform visual inspection of loaded ports before transferring the cartridge to the Analyzer.

Agilent Technologies Seahorse XFe96 Specifications