Eppendorf FemtoJet express - Manual (Page 24)

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The following are available:
• Adherent cells in a Petri dish,
• Injection liquid (purified, e.g. by centrifugation),
• Femtotips
®
/ Femtotips
®
II; other capillaries as an alternative
• Microloader,
• 0.5 – 10 µl pipette to take the Microloader,
• A universal capillary holder on the micromanipulator which has been preadjusted with a
Femtotip
® .
Filling the Femtotips
®
with the Microloader:
– Select function 0 "Capillary exchange" by pressing . Connect the pressure tube.
– Aspirate approximately 0.5 – 5.0 µl of centrifuged injection fluid with the Microloader
without touching the base of the vessel. Using the Microloader, fill the Femtotip
®
through
the rear opening, inserting the tip approximately up to the centre of the Femtotips
®
and
releasing the fluid there.
– Hold the Femtotip
®
vertically, loosen the cap by turning it and allow it to drop vertically.
– Screw the Femtotip
®
into the capillary holder.
– Complete Function 0 by pressing Compensation pressure is applied.
– Move the capillary into the cell medium (Petri dish) with the micro-manipulator.
– Focus the cells with the microscope, position the capillary slightly above the cells but do
not allow it to come into contact with the base of the Petri dish. Focus the capillary.
– Using the key, check that the capillary is not blocked. Either the fluid can be seen
flowing out or it is rendered visible by the fluorescent substance it entrains.
As a rule, any air bubbles present are blown out.
– Perform a sample injection using the preset parameters (e.g. p
i
~ 150 hPa, t
i
~ 0.5 s,
p
c
~ 50 hPa, corresponding to p
i
~ 2.18 psi, t
i
~ 0.5 s, p
c
~ 0.73 psi). Injection can be
monitored visually when the cell size changes by approx. 5 % to 10 % or via fluorescence.
If required, adapt parameters p
i
and/or t
i
.
3.5 The first injection
Mode of operation
3 Mode of operation
3
03_Bedienung_en.fm Seite 72 Freitag, 29. Juni 2007 2:49 14