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.
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