I've been watching some of your BB procedures and comparing it to the BB procedures from where I work. Thank you for this helps a lot in many ways since I live and work here in the Philippines and some or maybe most of the laboratories here don't have ready made reagents such as Known A1 and B cells, O cell suspension, etc. so we have to improvise on these stuffs.
Thank u for a good presentation.Is there any other method of preparing 3-5 percent saline apart from this method of comparing it with commercially prepared one?.The reason i ask is because some people practice where this commercially prepared one are not available.Thanks once again for sharing the knowledge
Don’t the tubes need to be caped or parafilmed before putting them in the centrifuge? Because I noticed in your video that you did not do so. Is that ok?
hey i have a few questions about this test. 1. Why need to have 3% cell suspension? 2. What is the purpose? 3. Different between using whole blood and 3% cells suspension.? -
+Oyleza Bell For optimal antigen/antibody reactions to occur, we need to avoid prozone and postzone. These involve an RBC suspension that is either too heavy or too light. Too much antigen or antibody can lead to false results.We don't use whole blood because every person may have a different RBC concentration. The essence of blood bank testing is consistency, so for that reason every step is controlled even the concentration of RBCs.
The suspension is made cause we should make sure that the concentration between the Ag and Ab of the test will be ballance. If you watched from the beginning of the video, he separated the plasma component to remove the Ab of the sample at first, it means that there's only RBC in that tube which means the Ag of that blood is much more than a whole blood.
What do you do if you have not reached the desired 3% suspension after putting in the saline? Do you just dump a little bit of the suspension then add more saline?
In our laboratory manual, it says we have to wash red cells at least 3 times. What does it mean? Please i really need your help. Our professor is being strict when we commit mistakes.
This is due to variability in procedures. Some facilities may require three washes and some only one for routine testing. You follow whatever your instructor, blood bank supervisor and/or standard operating procedure (SOP) tell you to do.
But why do we need to wash the cells in the first place? What are we washing away from the RBCs? What would happen if we didn’t wash the cells? Thank you 🙏🏾
The problem with using whole blood is that it still has the plasma in it, which is not what you want. You may have to wash your RBCs more than once. You should ask your instructor or blood bank supervisor.
The point of washing is to get rid of as much protein as possible, so washing a few times with isotonic saline will probably suffice. However, you really should consult your instructor or blood bank supervisor on this.
satish mani in my experience, we used the sample of the patients 3 days ago. the procedure of doing it, is like the red cell suspension. but first u have to do blood typing. so u can have blood type A and B positive. 3 drops of red cell from blood type a and b. then wash it using nss 3 to 4 times, decant, and then put nss to the last decant till it looks like a cherry color. and thats our known a and known b..
Hi Mohanad, if you are a student, doesn't your instructor have a procedure? If you work in a lab, doesn't it have a standard operating procedure (SOP) for a 5% suspension?
hmm what are the uses of RBC suspension in the laboratory? and how can centrifugation and decanting of supernatant affect the final concentration of the red cell suspension? pls help me, I really need the answers asap. thanks:)
In blood bank testing, we test plasma/serum and RBCs. For RBC testing, e.g., forward typing, we need a 3% RBC suspension from the patient before we can start the test. Another example of when you would need a 3% suspension is for a crossmatch. You take a segment from the donor unit and make a 3% suspension before you start crossmatch testing.Centrifugation and decanting aren't usually issues when preparing the 3% suspension. They are more of an issue for washing RBCs after an incubation, for example, during the indirect immunoglobulin test (IAT). When preparing the 3% RBC suspension, you have to be careful that you don't add too little or too much saline. To see if your 3% suspension is correct, compare it against a reagent bottle that has cells, e.g., A1 or B cells used for reverse typing.
Hi Hafsa, we make a 3-5% RBC suspension for two reasons. One is because we need to have consistent testing conditions. When we make a 3-5% suspension for all patients, we can say that the results will be more consistent and reliable. If we used whole blood, the percentage of RBCs could be very diffferent from patient to patient. The second reason is because whole blood contains plasma which may interfer with agglutination. To put it simply, plasma is sticky and may lead to false positives in agglutination.
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Thank you so much. We have retdems tommorow and this video reallyyy helped me recall all the procedures. Love from the Philippines ❤️
Thank you. I have worked with some very nice people from the Philippines. :)
Very informative and easy to understand, thank you for helping me study haha!
Thank you, Pat you rock!
Very well explained. Thank you so much . 💗💗
Wow very informative. Thnx for the video :)
thanks Pat
thank you patrick!:)
I've been watching some of your BB procedures and comparing it to the BB procedures from where I work. Thank you for this helps a lot in many ways since I live and work here in the Philippines and some or maybe most of the laboratories here don't have ready made reagents such as Known A1 and B cells, O cell suspension, etc. so we have to improvise on these stuffs.
I am glad they are of help to you. That's why I made them.
Really helpfull.
in our class we have to calculate the amt of red cell and nss to be added
Thanx alot
Good one
May I ask what do you call to the final color of the red cell suspension? Do you also call it tomato red or not?
Thank u for a good presentation.Is there any other method of preparing 3-5 percent saline apart from this method of comparing it with commercially prepared one?.The reason i ask is because some people practice where this commercially prepared one are not available.Thanks once again for sharing the knowledge
It may be possible to do a hematocrit on the suspension, which should be 3-5%, but I cannot guarantee in any way that these results will be accurate.
Patrick Tracy salam
I'm a new tech training in bloodbank now and we don't spin our suspensions, why do you? Thanks for the videos.
The purpose is to give the RBCs an initial wash, but not all facilities practice this.
Was your initial spin on pink top tube 60 seconds?
Yes
in anemic how many drops i can take please mention for making 3% cell suspension?
Satish,
When I make a 3% suspension, I compare it to a reagent bottle (see the video at 5:30).
But if we didn't have one in lab how we can do it
Don’t the tubes need to be caped or parafilmed before putting them in the centrifuge? Because I noticed in your video that you did not do so. Is that ok?
The serofuge is sealed, so the tubes don't need capping or parafilming.
When you centrifuge the blood in the EDTA how many minutes you used and the RPM of the machine?
3400 rpm for 5 minutes
hey i have a few questions about this test.
1. Why need to have 3% cell suspension?
2. What is the purpose?
3. Different between using whole blood and 3% cells suspension.?
-
+Oyleza Bell For optimal antigen/antibody reactions to occur, we need to avoid prozone and postzone. These involve an RBC suspension that is either too heavy or too light. Too much antigen or antibody can lead to false results.We don't use whole blood because every person may have a different RBC concentration. The essence of blood bank testing is consistency, so for that reason every step is controlled even the concentration of RBCs.
The suspension is made cause we should make sure that the concentration between the Ag and Ab of the test will be ballance. If you watched from the beginning of the video, he separated the plasma component to remove the Ab of the sample at first, it means that there's only RBC in that tube which means the Ag of that blood is much more than a whole blood.
@@ear_sthetic1221
Thank you sooooo much
What do you do if you have not reached the desired 3% suspension after putting in the saline? Do you just dump a little bit of the suspension then add more saline?
In our laboratory manual, it says we have to wash red cells at least 3 times. What does it mean? Please i really need your help. Our professor is being strict when we commit mistakes.
This is due to variability in procedures. Some facilities may require three washes and some only one for routine testing. You follow whatever your instructor, blood bank supervisor and/or standard operating procedure (SOP) tell you to do.
Thank you so much for your video. It really helped me. God bless!
But why do we need to wash the cells in the first place? What are we washing away from the RBCs?
What would happen if we didn’t wash the cells?
Thank you 🙏🏾
Do you have complete MLS course, so i can apply for to gain more experience.
No, just MLT.
Sir, why we use normal saline to make 3% rbc suspension. Why we do not use distilled water?
In order to maintain osmotic balance, we use isotonic saline. If we use water, the RBCs will burst.
Patrick Tracy thank you sir
Sir, at what rpm did you centrifuge?
3400-3500 rpm
Can you use whole blood from an unspun pink tube?
Thank you.
The problem with using whole blood is that it still has the plasma in it, which is not what you want. You may have to wash your RBCs more than once. You should ask your instructor or blood bank supervisor.
Thank you very much, Patrick!
So there would be no problem if it is washed a few times?
The point of washing is to get rid of as much protein as possible, so washing a few times with isotonic saline will probably suffice. However, you really should consult your instructor or blood bank supervisor on this.
Thanks again, Patrick!
Actually a fellow tech insisted on doing this method.
How to make pooled blood to do reverse grouping? please reply.
Hi Satish,
I really don't have any experience with that, so I cannot say.
satish mani in my experience, we used the sample of the patients 3 days ago. the procedure of doing it, is like the red cell suspension. but first u have to do blood typing. so u can have blood type A and B positive. 3 drops of red cell from blood type a and b. then wash it using nss 3 to 4 times, decant, and then put nss to the last decant till it looks like a cherry color. and thats our known a and known b..
Patrick Tracy plz ap ka number mil sakta he sir plz
May i ask what is the amount of the washed rbc?
I put in 1 drop of spun RBCs and 2-2.5 ml of 0.85% isotonic saline.
How long this suspension valid to use ?
I tried to find information about this but couldn't. Sorry, but can't give you an answer.
May I ask How can make 5% suspension ,what the procedure please?😊
Hi Mohanad, if you are a student, doesn't your instructor have a procedure? If you work in a lab, doesn't it have a standard operating procedure (SOP) for a 5% suspension?
hmm what are the uses of RBC suspension in the laboratory? and how can centrifugation and decanting of supernatant affect the final concentration of the red cell suspension?
pls help me, I really need the answers asap. thanks:)
In blood bank testing, we test plasma/serum and RBCs. For RBC testing, e.g., forward typing, we need a 3% RBC suspension from the patient before we can start the test. Another example of when you would need a 3% suspension is for a crossmatch. You take a segment from the donor unit and make a 3% suspension before you start crossmatch testing.Centrifugation and decanting aren't usually issues when preparing the 3% suspension. They are more of an issue for washing RBCs after an incubation, for example, during the indirect immunoglobulin test (IAT). When preparing the 3% RBC suspension, you have to be careful that you don't add too little or too much saline. To see if your 3% suspension is correct, compare it against a reagent bottle that has cells, e.g., A1 or B cells used for reverse typing.
Thank you patrick!:)
Merci beaucoup
pouvez-vous nous faire une vidéo sur l'hémolysine s'il vous plaît
You're welcome
Why we make rbc suspensions? Why not just use whole blood
Hi Hafsa, we make a 3-5% RBC suspension for two reasons. One is because we need to have consistent testing conditions. When we make a 3-5% suspension for all patients, we can say that the results will be more consistent and reliable. If we used whole blood, the percentage of RBCs could be very diffferent from patient to patient. The second reason is because whole blood contains plasma which may interfer with agglutination. To put it simply, plasma is sticky and may lead to false positives in agglutination.