Lonza 4D-Nucleofector - LV Unit (Scalable Volume): Mounting Cartridge

4D-Nucleofector
®
Manual | 27
2.12.2.3 Mounting an LV Nucleocuvette
®
Catridge
1. Inset the LV Nucleocuvette
®
Catridge into the slot.
Make sure that upper and lower snap mechanism
audibly engages to allow for proper contacting.
2. Attach the venting tube with the filter (upper left of
the catridge) to the male Luer connector on the
front plate (Figure 2.37, A).
3. If you stated defining your experiment setup already
(see 2.12.2.1), the software can suppot you in inset-
ing the tubes into the pinch valves by automatic
opening of the valves. On the right side of the experi-
ment screen you can select “Yes” if you want to inset
the tubes of the LV Nucleocuvette
®
Catridge into
the pinch valves now (Figure 2.37, B). Alternatively,
you can open the pinch valves manually by pushing
the black button on the valves.
4. Upon pressing “Yes” the pinch valves will automati-
cally open for 30 seconds allowing to conveniently
inset the tubes into the three pinch valves (Figure
2.37, C).
NOTE: The open time of the pinch valves is limited to 30
seconds to avoid overloading of the electronics.
5. Inset the tubes into the three pinch valves (Figure
2.37, D). Ensure that they are inseted completely
(Figure 2.37, E). After 30 seconds, the pinch valves will
close automatically. In case you did not finalize inser-
tion within 30 seconds, you may re-open the valves
by pressing “YES” again or manually (see “Note”
above).
6. Inset the T connector on the left hand side of the
catridge into the T connector holder (Figure 2.37, F,
left).
7. Inset the two larger diameter tubes into the black
holders (figure 2.37, F, right). Ensure the tubes are
inseted completely into the black holders. The lower
holder comprises a light sensor for liquid detection in
the lower tubing line.
8. Inset the upper tube into the upper pump and close
the flap (Figure 2.37,G). When closing the flap, gently
hold the tube without pulling it.
9. Repeat with the lower tube and the lower peristaltic
pump (optional, if working with 2 input reservoirs).
10. Fill the output reservoir with medium (for detailed
recommendations, please refer to cell type-specific
protocol) and mount it into a 4D-Nucleofector
®
LV
Reservoir Rack (Figure 2.37, H).
2
11. Remove the red cap from the Spiros connector on
the outlet tube of the catridge and connect it to
the swabbable injection pot of the output reservoir
(or any other reservoir with female Luer connector,
figure 2.37, I). Alternatively, when working with the
weldable LV catridge version, you may establish the
connection by welding.
12. Before continuing check setup status: At this point
steps 1–6 and 8 of the arrangement shown in figure
2.31 should be done.
13. Continue with sample loading (see 2.12.2.4).
2.12.2.4 Loading Samples
1. Prepare cell suspension under the sterile hood (for
detailed recommendations, please refer to cell type-
specific protocol).
2. Fill a defined volume of cells and substrate into the
input reservoir(s) mounted on a 4D-Nucleofector
®
LV
Reservoir Rack.
3. Place the rack with the cell suspension reservoir on
a magnetic stirring platform to avoid cell sedimenta-
tion when working with larger volumes. Stat stirring
the cell suspension at ~300 rpm. Ensure that magnet
is truly stirring.
4. Remove red caps from the Spiros connectors on the
inlet tubes of the catridge and the blue caps from
the swabbable injection pot of the output reservoirs
and connect both (Figure 2.37, G). Alternatively, when
working with the weldable LV catridge version, you
may establish the connection by welding.
5. The system is now fully assembled. Check correct as-
sembly as shown in figure 2.38.
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