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Dear
Many thanks to those who respond my question! The
information you provided are very helpful
I summarize all the responses here
Thank you again
Best regards and Merry Christmas.
Simona
=======================================================================================================
ORIGINAL QUESTION
Hi all,
I have a problem with Fill and nozzle flush on
Facs Vantage SE + turbo optional
I work at 20 PSI and when the nozzle is clogged I
put on fill and I push the bottom nozzle flush,
but the stream go to back very slowly and is not
enough to clean the nozzle.
This happen also when the nozzle is not clogged!!
Any comments are welcome
Many thanks in advance
Simona
======================================================================================================
RESPONSES
I have found that if it is quite bad I can
usually unclog the nozzle by filling a syringe
with a small tube on it that fits on the nozzle
tight with bleach. Then I turn the system to
Fill, release the pressure from the tank, put the
tube on the nozzle, push nozzle flush and squirt
in some bleach at the same time. It is much
easier than taking off the nozzle. If it is not
very bad, sometimes putting a small cap or cut
transfer pipet end with bleach up to the nozzle
while nozzle flushing works better than just
flushing alone.
-will
William Schott
Flow Cytometrist
The Jackson Laboratory
600 Main St.
Bar Harbor, ME 04609
Phone: (207) 288-6192
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Simona,
Often times when you think the nozzle is clear of "junk" it is still
dirty. Try removing the nozzle and sonicate for 10 minutes in 10%
bleach. Rinse well with deionized water. I dry the nozzle with 70%
alcohol before replacing back onto position. obviously, you will be
required to align the 488 laser at this point. Hope this helps.
Daniel Marmer, M.S., MT(ASCP)
Manager, Core Flow Cytomerty Facility
Cincinnati Children's Hospital Med Center
Division of Hematology/Oncology
3333 Burnet Ave, Bldg R, ML 7015
Cincinnati, Ohio 45229
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Hello Simona-
I will explain what I think you're describing,
and provide two ideas as feedback.
First, it sounds like the stream does not stop
flowing quickly, when you reverse flow. If this
is the case, there is likely some air trapped in
the nozzle, or in the sheath lines. If your
nozzle is not clogged (plugged), the easiest way
to treat this, is to aspirate 70% ethanol through
the nozzle orifice.
Second, whenever I prepare to sort, or to unclog
a nozzle, I aspirate 20%-50% bleach through the
nozzle. While aspiration is happening, I remove
the bleach, to allow air to be taken into the
nozzle, then put the bleach back. I do this
repeatedly, after which I allow the normal sheath
flow through the nozzle for at least 5-10
minutes. This clears most clogs (debris in
orifice), and also sterilized this area (nozzle)
which is very susceptible to contamination, as it
is often open to the air. Be sure to rinse the
nozzle area with large amounts of diH2O, to
remove all bleach.
I hope this helps. Please contact me if you need
further explanation, or if I have misunderstood
you!
Buona sera!
Andrew
Andrew Beernink
Research Scientist
Manager, Flow Cytometry Facility
Novasite Pharmaceuticals, Inc.
11025 Roselle St.
San Diego, CA 92121
(858) 875-8584
(858) 875-8501 fax
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Hello Simona,
Have you checked your vaccuum? Is it high enough
to aspirate and flush the line?
Also, maybe there is a clogg in the vaccuum line
or a leak on the waste tank. It can possibly
cause a problem while trying to flush the nozzle.
Good luck.
Eric Massicotte
Responsable, Service de cytométrie en flux
Institut de Recherches Cliniques de Montréal (IRCM)
110 ave. des Pins Ouest
Montréal, Québec
Canada, H2W 1R7
Téléphone: (514) 987-5724
Télécopieur: (514) 987-5736
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Hi Simona,
You may have to ask your service engineer to replace the solenoid valve
which controls the nozzle flush. Another thing may be the problem may
be the vacuum pump. You may not be getting enough vacuum to the
instrument. I suspect, though, that the solenoid valve is the problem.
Keyvan
----
Keyvan Keyvanfar
Flow Cytometry Laboratory
Hematology Branch, NHLBI, NIH
Building CRC RM 3E-5217
10 Center DR. MSC 1202
Bethesda, MD. 20892-1202
Phone#: 301-402-0277
Fax #: 301-496-8396
keyvanfk@nhlbi.nih.gov
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Hi Simona,
Check your vacuum source, and all the fluidics between your vacuum
source and the nozzle.
Good luck,
Eric
Eric Van Buren <eric.vanburen@wayne.edu>
Manager, Flow Cytometry Core Facility
Karmanos Cancer Institute
Detroit, Michigan, USA
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At low pressures (e.g. below 20psi) this occurs, although on my DiVa it is
usually only below about 12psi. I believe it has to with the amount of vacuum
applied at the waste tank. My house vacuum is about 9psi on a good day (I have
extra ports on my tanks for measuring with a digital manometer -- long story).
Here is a convoluted work-around that I have which was actually the spurious
effect of another fix: One day my DiVa decided to backflow waste through the
collector when I was in nozzle flush. It started suddenly and I still haven't
discovered the root cause. As a repair, I routed a second vacuum line from my
waste tank to the nozzle flush vacuum line after it comes out of the solenoid
behind the fluidics knobs (can't remember offhand what number solenoid, but
I'll look it up for any interested parties). This is routed to a quick-connect
at the tank, so I am now using all four ports on my tank (normal waste, nozzle
wastem vacuum source, and measurement port). By doing this, I have completely
divorced the collector waste from the nozzle flush waste so there is no
possibility of backflow unless my entire waste tank fills up.
Now, the workaround that I discovered with the low pressure sorts is that when
I nozzle flush, if i pinch the new waste hose just the right amount, it
attenuates the vacuum applied and I get a proper fill stream.
Sounds easy, doesn't it?
-Christopher Bare
Senior Flow Cytometry Specialist
Sloan-Kettering Institute
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Simona
First, check that you have full vacuum to the
machine from the wall. The house vacuum lines
corrode, due to the bleach vapors from your waste
tank.
I've also found that the plastic vacuum lines in
the machine can become clogged over time. You can
try opening the control panel and use clamps to
close the line from the stream waste collector to
get more vacuum to the nozzle. I had to flush
(several times!) the whole pathway from the waste
collector to the tank with detergent. I forced
detergent BOTH ways to flush the tubing, and I
saw that it was indeed quite full of junk. The
little fittings can become clogged easily.
---Dennis
Dennis J. Young
Flow Cytometry Shared Resource
Moores UCSD Cancer Center
Room 2314 Bay 2Q
3855 Health Sciences Drive #0803
La Jolla, CA 92093-0803
Mail:<<mailto:djyoung@ucsd.edu>>
WWW:<<http://cancer.ucsd.edu/flow/>>
Telephone:(858) 822-0407 FAX: (858) 822-0403
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Hi Simona
We use a FACSVantage as well, and have frequently had the same problem.
Here are some ideas:
1) the commonest problem is clogging, so try
sonicating the nozzle for about 1 minute
2) the other problem may be a leak in the vacuum
seal, so the fluid cannot be drawn up. This may
be due to a leaky O-ring.
3) the other problem may be alignment of the
stream with the laser, so that most of the
'events' are not registered with the laser. The
problem with FACSVantage is that the laser is
'free standing' and so it needs alignment at
least every day. We had a problem that we had
not tightened the stream unit, and so the stream
would go out of line very quickly, and then the
count fell.
The BD people who make the machine are very helpful if you email them.
Good luck,
Hemali
MD Student
Warwick University, UK
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Dear Simona,
I experienced a similar problem with our
FACSVantage SE (without turbo option). What I
discovered was that the valve in the vacuum/waste
line from the nozzle holder had a stuck solenoid
plunger. This is the third valve from the top or
from the front with the valve panel pulled down.
At first I thought that the solenoid had burned
out but when I removed the valve head it was
apparent that the piston had salt crystal
build-up that had made it stick. You may try
rapping the solenoid body to jar the plunger
loose but I would recommend opening the unit and
cleaning the plunger.
Try the same operations that you describe with
the nozzle removed. If the flow stops immediately
when you press FLUSH, then it is probably the
same problem that I described above. The
continued flow when the nozzle is in place is
because of air bubbles in the nozzle that are
expanding and continuing to press fluid out the
nozzle even though the sheath valve is shut off.
Best Regards,
Brad Hernlem
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Hi Simona,
I'd try sonicating the nozzle for a minute. If this fails then
maybe one of your fluidic solenoid valves is defective....Ken -
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--
Simona Ronzoni
Flow Cytometry and Imaging Core
Experimental Oncology Department
European Institute of Oncology
via Ripamonti, 435 20141 Milan Italy
Phone: ++39-02-57489880 Fax: ++39-02-57489851
e-mail: simona.ronzoni@ifom-ieo-campus.it
http://www.ifom-ieo-campus.it
http://imaging.altervista.org
