keebie81 Posted May 30, 2012 Share Posted May 30, 2012 Forgive me if this has been covered already. I tried looking and did not see it anywhere.I work in the biomed dept and got a work order for the cell washer washing away all the cells. We have a Helmer UltraCW. I have spoke with tech support and we have gone over all the settings and they are correct. But it still washes away the cells, they even sent me a loaner machine and with same settings i get the same results.We are using Panocell 2-4% Reagent Red Blood Cells for testing. The way lab showed me to test is to put a drop in each of the 6 tubes, run the cycle and see if there is a cell button formed at the bottom. Everytime I do this I end up with nothing. But I have noticed if I press the check button after each step and look there is a button cell at the end of the 4cycles. I am looking for how other labs use this device for testing. Maybe our method is wrong. The operator manual isnt much help except for setting the machine up. Link to comment Share on other sites More sharing options...
SMILLER Posted May 30, 2012 Share Posted May 30, 2012 We have a Helmer CW but I am pretty sure we have never had this problem. I am not sure if there is a setting that can be used that would cause this sort of thing even if you wanted it to. Make sure that the rotor is fully seated. The thing is spinning between cycles, right? The cell button should stay in the bottom of the tube when they are tipped and dumped after spinning. Has anything else changed recently that might have caused this (like new test tubes or something)? Other than that, you have to call Helmer back and ask for more help. They are responsible to make sure that thier products are being used effectively. You should be able to find someone who can explain what is being done wrong.Good Luck, Scott Link to comment Share on other sites More sharing options...
keebie81 Posted May 30, 2012 Author Share Posted May 30, 2012 It is spinning, the rpms are all good. It seems to have a cell button but during the decant its gets lost. If i put the tubes in a centrifuge after a button forms Link to comment Share on other sites More sharing options...
John C. Staley Posted May 30, 2012 Share Posted May 30, 2012 Just a thought to rule out. Is the cell washer hooked up to saline or H2O? I know this is a "silly" question but I have seen it happen. The cells are not washing out they are hemolyzing. Just need to cover all possibilities. Link to comment Share on other sites More sharing options...
keebie81 Posted May 30, 2012 Author Share Posted May 30, 2012 Just a thought to rule out. Is the cell washer hooked up to saline or H2O? I know this is a "silly" question but I have seen it happen. The cells are not washing out they are hemolyzing. Just need to cover all possibilities. I am using a SP Certified Blood Bank Saline cube from Cardinal Health Link to comment Share on other sites More sharing options...
Malcolm Needs ☆ Posted May 30, 2012 Share Posted May 30, 2012 Good point John (as ever).Joyce Poole is for ever telling me not to reinvent the ABO Blood Group System when, for example, I test a red cell, supposedly Lan negatvie, get it Lan positive, and then realise it is group A, and the anti-Lan I'm using is only suitable to test group O red cells. Grrrrrrrrrrrrrrrrrrrrrrrrr!!!!!!!!!!!!!!!!! Link to comment Share on other sites More sharing options...
Mabel Adams Posted May 30, 2012 Share Posted May 30, 2012 It sounds like when the cells are mixed up into the wash saline, they are being decanted out. Once cell button is produced in the bottom of the tube by centrifugation, it is pretty hard to dislodge all of it with the normal decanting process. It is much easier to lose all of the cells while they are suspended in the saline. Overfilling the tubes during the fill cycle can probably do this. I will have to look at our Helmer. I am more familiar with the decant mechanisms of our old Dade and Sorval cell washers. If the saline is really saline (and only saline) then that rules out water or bleach or anything else that might hemolyze the red cells. Has it all been the same cube of saline? Was it new when the problem started. Maybe it was manufactured with the wrong amount of salt? Test the specific gravity of the saline in the urinalysis dept and compare with what it should be (by calculation) or what a cube from another lot is. Link to comment Share on other sites More sharing options...
keebie81 Posted May 31, 2012 Author Share Posted May 31, 2012 It sounds like when the cells are mixed up into the wash saline, they are being decanted out. Once cell button is produced in the bottom of the tube by centrifugation, it is pretty hard to dislodge all of it with the normal decanting process. It is much easier to lose all of the cells while they are suspended in the saline. Overfilling the tubes during the fill cycle can probably do this. I will have to look at our Helmer. I am more familiar with the decant mechanisms of our old Dade and Sorval cell washers. If the saline is really saline (and only saline) then that rules out water or bleach or anything else that might hemolyze the red cells. Has it all been the same cube of saline? Was it new when the problem started. Maybe it was manufactured with the wrong amount of salt? Test the specific gravity of the saline in the urinalysis dept and compare with what it should be (by calculation) or what a cube from another lot is.Ive gone through a few different saline cubes, the lot numbers have been different. The tubes are 12x75mm and filled with 4.7ml salineThe wash(rpm) is set to 3500, Twash 0:35, decant(rpm) 425 Link to comment Share on other sites More sharing options...
Barbarakym Posted May 31, 2012 Share Posted May 31, 2012 My first thought when this happens is the tubes are overfilling. There is an adjustment on the tubing which you need to do as PM (check volumn dispensed). If move than allowable for the cell washer, the cells are filling up too much and during mix cycle more and more cells get decanted as they 'spill over'. This is the first thing I check and maybe you did, but I don't see it mentioned. Link to comment Share on other sites More sharing options...
keebie81 Posted May 31, 2012 Author Share Posted May 31, 2012 My first thought when this happens is the tubes are overfilling. There is an adjustment on the tubing which you need to do as PM (check volumn dispensed). If move than allowable for the cell washer, the cells are filling up too much and during mix cycle more and more cells get decanted as they 'spill over'. This is the first thing I check and maybe you did, but I don't see it mentioned.Yes i checked that, measured 56.4mL. This is the same value shown on display. I also measured the volume of the 12x75mm tube, it was around 4.5mL. Link to comment Share on other sites More sharing options...
Barbarakym Posted May 31, 2012 Share Posted May 31, 2012 Yes i checked that, measured 56.4mL. This is the same value shown on display. I also measured the volume of the 12x75mm tube, it was around 4.5mL.You have cells after 4 cycles. I just really still think overfilling is the issue. Why not as experiment adjust so volume is 52-55... Just test it and see. Good luck. Link to comment Share on other sites More sharing options...
mkeepes Posted June 1, 2012 Share Posted June 1, 2012 Yes i checked that, measured 56.4mL. This is the same value shown on display. I also measured the volume of the 12x75mm tube, it was around 4.5mL.You might check to see if the "global setting" for Spin Decant M is on or off. I think it needs to be off. To get to the global settings press the left arrow until they appear and then press the left arrow to toggle through all of the settings, when finished press Start Wash to save or exit. Link to comment Share on other sites More sharing options...
keebie81 Posted June 4, 2012 Author Share Posted June 4, 2012 You might check to see if the "global setting" for Spin Decant M is on or off. I think it needs to be off. To get to the global settings press the left arrow until they appear and then press the left arrow to toggle through all of the settings, when finished press Start Wash to save or exit.Thanks, it was to on. It seems to be doing better right now. After the spin step i can see a button and it looks good to me. But still after the decant the button breaks apart and looks runny. I will try attaching an image in a bit. Link to comment Share on other sites More sharing options...
keebie81 Posted June 4, 2012 Author Share Posted June 4, 2012 Here is an attached image of after spin and after decant Link to comment Share on other sites More sharing options...
keebie81 Posted June 4, 2012 Author Share Posted June 4, 2012 I turned the decant speed down quite a bit and seem to be getting a decent result. At the end i have about 1 drop of saline in the tube Is this an acceptable amount? The decant is at 368rpm Link to comment Share on other sites More sharing options...
mkeepes Posted June 9, 2012 Share Posted June 9, 2012 Here is an attached image of after spin and after decantI think the pictures of the tubes look as expected after centrifugation and after decanting. The cell button won't be a tight button after decanting and there should be a small amount of saline present with it. I am setting up our first Helmer cell washer (always used IEC Centra W or Sorvall before) and have played with a lot of settings to see what worked best for us and I ended up back at the recommended settings (on the blue quick reference guide) for almost everything except I added a few more shakes (agitation) between washes because the cell button wasn't completely resuspended in a few tubes after addition of saline prior to centrifugation during wash cycle. I have our decant speed set to 435. I think the default is 440. Link to comment Share on other sites More sharing options...
mkeepes Posted June 9, 2012 Share Posted June 9, 2012 From what I understand if the button is too wet, increase the decant speed and if the cells stream up the side of the tube, decrease the decant speed. Good luck! Link to comment Share on other sites More sharing options...
Mabel Adams Posted June 10, 2012 Share Posted June 10, 2012 the instrument specs might list the expected residual saline volume. Link to comment Share on other sites More sharing options...
KathleenSL Posted June 15, 2012 Share Posted June 15, 2012 We use 10 x 75 mm test tubes so our saline volumes are different. I have attached the settings that we use for our various programs. Link to comment Share on other sites More sharing options...
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