26 | 4D-Nucleofector
®
Manual
2.12.2.2 Defining a new experiment
Following vessel type selection (see chapter 2.8.2) you can
now either select a predefined experiment (see chapter
2.8.4 or create a new experiment).
If you want to create a new Experiment, tap in the field “Ex-
periment name” and enter a specific experiment name or
accept the suggested name generated with date and time
stamp. Press “Submit” to get to Experiment setup screen
(Figure 2.32).
1. Enter Cell Type Code (optional)
Press the field “Cell type code” to choose predefined
Nucleofection
®
Conditions from a cell type list (Figure
2.33). A list with all available Cell type codes (defined
by Lonza or custom) will appear. Select the code that
shall be used by scrolling through the menu or stat
typing in the search field . To confirm your selection
press “Apply”. If you pick a preset Cell type codes the
next two steps do not apply.
NOTE: Instead of defining solution and program code via
the Cell type code, both parameters can also be selected
manually (step 2 and 3), e.g. in case no predefined Cell type
code is available. For adding new Cell type code, please
refer to chapter 2.13.2.3.
2. Enter Pulse code
Modify pulse code if necessary by pressing the “Pulse
code” field. A keyboard will appear, enabling you to
change settings. To confirm your choice, click “Apply”.
3. Enter Solution
The solution code can be modified via selecting the
“Solution” field and click on the desired solution. Then
press “Apply” to confirm the selection.
4. Enter the volume you plan to process
Press on the field “Volume” (Cell suspension volume)
to define the planned volume of your cell suspension
(up to 20 mL are possible).
a. If you work with one reservoir (or bag) containing a
premixed suspension of cells and substrate, enter
the total volume of the cell suspension you want to
process.
b. In case you work with two reservoirs (or bags) to
keep cells and substrate separate, tick the box
“Substrate separate”. A second volume field will
show up for “Substrate” to define the volume of
substrate independently. In addition a field will ap-
pear named “Bubble detection”:
• If bubble detection is de-activated (default setting),
the liquid sensor is only used for priming of the
substrate line (detection of liquid stat). Afterwards
the system would ignore any air fractions within the
substrate feed and continue processing until the
end of liquid according to volume entry is reached or
if multiple errors occur that point to missing liquid.
However, the air fraction may lead to an increased
error frequency. In addition larger fractions without
substrate may lead to a decrease in transfection ef-
ficiency, as the sample is pulsed without substrate.
• If bubble detection is activated, the system will inter-
rupt the pulsing process in case the substrate line
contains a larger bubble (>7 mm). A message will
appear and the user can decide whether the process
shall be continued or aboted. In case “continue” is
selected, the system will continue the process like
in the “de-activated” setting, i.e. it would ignore any
futher air fractions in the substrate feed and con-
tinue processing until the end of liquid according to
volume entry is reached or if multiple errors occur
that point to missing liquid.
NOTE: The maximum total volume that can be entered is
20 mL and the ratio between Cell suspension and Substrate
volume should be between 1:1 (e.g. 10 mL cell suspension
and 10 mL substrate) and 9:1 (e.g. 18 mL cell suspension and
2 mL substrate)
5. Save your experiment (optional)
At this stage, you can save your defined experiment
or only the pulse-solution code combination (as
custom Cell type code) for future use by pressing the
“Save” button. Select whether the Cell type code or
the experiment shall be saved. A keyboard will appear
allowing you to define a name (max. length: 26 char-
acters) for either the Cell type code or the experi-
ment depending on your previous selection.
6. Continue with mounting the LV Nucleocuvette
®
Catridge (see 2.12.2.3)
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