Transcript FACSAria Basic Parts

FACSAria
The basic parts
Overview
Basic Parts of the FACS Aria
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.
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.
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.
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.
Basic Parts of the FACS Aria
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!).
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!
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...
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.
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)!
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.
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.
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
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!
Basic Parts of the FACS Aria
Exchanging the sample lines will be explained during
the trainings course.
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.
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.
Basic Parts of the FACS Aria
The three (depending on the configuration) laser
intercepts are focussed onto three distinct collection
fibers.
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.
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.
Basic Parts of the FACS Aria
Schematics of the
fluidics. After
analysis the cells
are either sorted or
directed into the
waste.
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.
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!
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.
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
4°C! Additionally, the chamber needs 30 minutes to
reach the desired temperature!
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.
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.