Eppendorf FemtoJet - Manual (Page 26)

24
3 Mode of operation
3.5 The first injection
The following are available:
● Adherent cells in a Petri dish (possibly on CELLocates to observe
specific cells after injection),
● 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 injection 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
.
03_Bedien_en.fm Seite 24 Mittwoch, 27. September 2006 5:12 17