Oroboros Instruments O2k Manual page 16

MiPNet15.03
4.5.2. Calibration range
4.5.3. ISE-calibration solution
4.5.4. TPP +
Initial concentration:
Oroboros Instruments
O2k-MultiSensor-ISE
The experimental TPP
the limit of detection and below the inhibitory
concentration (O2k-Procedure MiPNet14.05). Decide
on a concentration range and steps to be used for
calibration, e.g. 0.7 µM to 1.5 µM TPPCl, in 5 steps:
0.7, 0.9, 1.1, 1.3, and 1.5 µM, respectively. The
electrode should be allowed to stabilize at the lowest
calibration concentration.
may be filled with medium already containing a
minimum analyte concentration.
The ionic background of the solution should be close to
the experimental medium. The best option is to use
experimental medium directly.
MiR06 as a medium, 100 mM KCl solution is sufficient
for TPP + calibration, thus reducing the use of the more
viscose MiR06
syringes. The analyte concentration in the calibration
solution should allow for injection volumes small
enough not to create major disturbances, but large
enough to allow for precise injections. In our example
a 100 mM KCl solution containing either 0.1 or 1 mM
TPPCl present good choices when using the TIP2k
(which allows precise handling of very small volumes).
When the calibration
injections, a 0.1 mM solution is used.
calibration with the TIP2k
Fill the O2k-chambers with medium and close the
chamber with electrodes inserted as described above.
Fill the TIP syringes with the calibration solution and
insert the TIP needles into the chambers.
100 mM KCl. A first injection of 1.4 µl into the 2 mL
O2k-Chamber increases the chamber concentration by
0.7 µM TPP + . This is performed with a TIP2k program:
Line Mode Delay
S
1 D 1
Set the pX gain to 20 and allow the ISE-signal to
stabilize. The time derivative (slope) of the raw pX
signal (Section
. Drift is higher at extremely low (especially zero)
analyte concentration.
+ concentration should be above
Alternatively, the chamber
medium, particularly
is performed
Use calibration solution 1 mM TPPCl in
Volume Flow
µl µl/s
1.4 40
5.3) should be in the range ±0.04 mV/s
Mitochondria and cell research
When working with
with TIP2k
by manual
Interval Cycles
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1 1
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