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4.2.3 Spinning
1
Make sure the operation mode of the ultracentrifuge is ZONAT. And set speed of
revolution 3,000 rpm.
Then, turn on the start switch.
CAUTION :
If the lubricating unit becomes defective during operation, the lubricating unit makes beep
tones and an alarm signal. In this case, solve the problem quickly and clear the alarms.
(Refer to Section 3.3 for details.)
2
Loading
While spinning at 3,000 rpm, inject from the
outer connector overlay solution, cushion solution,
etc. from lighter ones.
Piping and clamp positions at this time are
shown on the right.
¡
In case of density gradient centrifugation,
inject 100-150 ml of overlay solution and then
330-280 ml of density gradient solution from
the lighter one, at a flow rate of 30-50 ml/min.
When the rotor is filled with density gradient
solution, the injection pressure becomes more
than 0.5 kgf/cm
2
.
This is not abnormal and can be corrected by
filling with the overlay solution in the
subsequent stage.
3
After the density gradient solution, etc. are
injected, change the piping as illustrated, and
allow overlay solution from the central connector
at a flow rate of about 30 ml/min.
When overlay solution overflows from the
central connector, the density gradient solution
has been injected completely.
¡
When changing the piping clamps, take care
not to allow air into the central connector, if air
is introduced, flow of solutions might stop in
the rotor or seal attachment at a high speed
spinning. To purge the air in the rotor and seal
attachment change the flow course between 2
and 3, 2 or 3 times repeatedly. Particularly, if
air enters the central connector, a trouble will
possibly take place. When changing piping in
the figure on the right, move the clamp in the
order of (A) and (B).
Central connector
Overall inlet
Clamp
Clamp
Outer connector
Central connector
Outer connector
Inlet for overlay solution and
density gradient solution
(B)
(A)
