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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ------------------------------------------------------------------------------------------------------------------------ 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 ----------------------------------------------------------------------------------------------------------------------- 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 - ------------------------------------------------------------------------------------------------------------------------ -- 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.orgReceived on Fri Dec 23 20:38:00 2005
This archive was generated by hypermail 2.1.8 : Sat Jan 14 2006 - 22:04:01 EST
