FACSAria The basic parts

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FACSAriaThe basic parts
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Overview
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Basic Parts of the FACS Aria
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Basic Parts of the FACS Aria

• Power switches
• Located at the left side
• Main Power turns on and off the Aria and the
  fluidics cart (via umbilical cable).
• The three additional switches turn on and off the
  power for the three different laser lines. Lasers
  can be turned off if not required for an
  experiment.

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Lasers- On and Off

• The Manual recommends to keep the lasers off
  until the stream is on to protect the "dry"
  cuvette from direct laser light.
• However, if you do this, the instrument will
  disconnect from the software most of the times
  you turn the lasers on....
• Solution Start with all required lasers on! Keep
  the front cover open to protect the cuvette until
  the stream is turned on.

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Basic Parts of the FACS Aria
The circuit breaker needs only to be switched if
there have been power surges in the laboratory.
Switch first OFF, then back ON again. Do not use
it as an additional power switch.
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Basic Parts of the FACS Aria
The fluidics cart provides air pressure and
vacuum. It harbours the sheath and waste tanks
and provides additional tanks for ethanol, water
and cleaning solution.
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Basic Parts of the FACS Aria
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Basic Parts of the FACS Aria
There should be no air bubbles in the filters. On
new instruments or after filter changes, inspect
the filters. If an air cushion is visible, vent
the filter through the bleeder valve (power must
be on, stream can be off!).
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Basic Parts of the FACS Aria
All tanks are pressure free. Thus, they can be
opened and refilled online during the
instrument is running. However, disconnecting the
fluidics lines from the ports requires to prime
the lines after reconnection. Disconnect the
lines only if you wish to clean the tanks. Do
not disconnect the lines while the stream is on,
because priming of the lines requires the stream
to be off!
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Basic Parts of the FACS Aria
The fill level of the tanks is shown with small
icons in the software. An error message will pop
up to remind you to fill/empty the corresponding
tank. Do not ignore this message! The sheath tank
may run dry, hampering your sorts. The waste tank
may overflow, requiring you to clean the lab...
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Basic Parts of the FACS Aria
The umbilical connection to the fluidics cart is
properly configured during installation. The user
can connect the temperature control and the AMS
to the depicted ports.
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Basic Parts of the FACS Aria
The connection from the fluidics cart to the main
instrument is configured during installation.
Users may supply house air into the depicted
port and switch off the internal compressor. The
house air must at least supply 6 bar (90 psi)!
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Basic Parts of the FACS Aria
The sample port is software controlled. The
sample chamber can be illuminated to check the
fluidics level of the sample.
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Basic Parts of the FACS Aria

• The system is delivered with different tube
  holders
• for 15 ml tubes
• for 12x75 mm standard tubes
• for 1 ml microtubes
• These holders are inserted into the sample port.
  Never insert tubes without the appropriate
  holder, as the sample station may become damaged.

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Basic Parts of the FACS Aria

• The system is provided with three different
  collection tube holders
• for 15 ml tubes only two way sorting is possible
  (not enough deflection for four tubes)
• for 12x75mm standard tubes
• for 1 ml microtubes

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Basic Parts of the FACS Aria
During operation of the sample port a red
emergency stop button is lit. In case anything is
hampering the operation of the chamber loading
mechanics, pressing this button will

• Unload the tube
• Stop the ACDU moving
• Shut off the stream
• Turn off the plate voltage
• Protect the sample by closing the waste drawer
• Put a warning message on the screen
• Thereafter the button can be reset by turning it
  to the left.
• Use with caution! The system has to be started up
  completely again!

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Basic Parts of the FACS Aria
Exchanging the sample lines will be explained
during the trainings course.
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Basic Parts of the FACS Aria
Steering optics direct the laser light to the
cuvette. The droplet breakoff point is observed
by a dedicated camera.
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Basic Parts of the FACS Aria
Schematic drawing of the interrogation area. The
cells are focussed in the flow cell, pass the
interrogation point and thereafter enter the
nozzle.
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Basic Parts of the FACS Aria
The three (depending on the configuration) laser
intercepts are focussed onto three distinct
collection fibers.
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Basic Parts of the FACS Aria
Each fiber leads to a dedicated detection block
based on the Octagon principle. Each signal
passes only one mirror. All mirrors are longpass
and thus reflect light with lower wavelenghts.
Due to this, an octagon can detect eight signals,
seven colors plus the blue side scatter.
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Basic Parts of the FACS Aria
Signals from the red and violet lasers are
directed to Trigons. At the moment there a few
combinations for more than three colors fomr the
red and violet lasers.
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Basic Parts of the FACS Aria
Schematics of the fluidics. After analysis the
cells are either sorted or directed into the
waste.
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Basic Parts of the FACS Aria
The sort chamber contains the plates and is
closed during normal operation (Aerosol
protection). The position of the red AccuDrop
laser can be adjusted by a micrometer dial.
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Basic Parts of the FACS Aria
Whenever the high voltage for the plates is on, a
red warning light is lit. Take care when opening
the sort chamber, as the plate voltage is about
6000 Volt!
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Basic Parts of the FACS Aria
The sort drawer protecs the samples and is only
opened during sorts. The FACSAria detects clogs
and closes the drawer automatically to protect
your sorted samples. You can of course open and
close it from the software manually.
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Basic Parts of the FACS Aria
The sample chamber can be set to a specified
temperature to keep the sample cold or warm. It
can only keep the sample at a given level, do not
put a warm sample in and expect it to cool down
to 4C! Additionally, the chamber needs 30
minutes to reach the desired temperature!
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Basic Parts of the FACS Aria

• There is a bug in the sample cooling
• If you quit the software and start it the next
  morning, the sample cooling is still set to the
  desired temperature in the menu, however, the
  unit is NOT working. You have to turn it off and
  then select the desired temperature again in the
  software! You can feel the breeze outside of the
  exhaust fans.

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Basic Parts of the FACS Aria
The system has an inbuilt agitator to keep cells
in suspension during sorting. However, bare in
mind that this only keeps cells suspended, it is
not a Vortex. Carefully resuspend all samples
before introducing them to the sample chamber.