hello...i took a blood sample with EDTA,0.9%NACL,glucose,buffer...inspite of this..after some time heamoglobin is getting leaked out after centrifuge...without any external sterss..what more should i add/do to prevent this leakage..and keep erythrocytes suspended??
You may need to reduce the speed of the centrifuge. Spinning the cell samples too hard can cause lysis, especially if the erythrocytes are stressed or damaged in any other way.
Dear Jyothi Atmakuri ,
In fact, media used for RBC conservation before transfusion ie are important, so allowing to preserve sample at 4°C until 30-40 days.
These can be also usefull for you in research, because these media avoid hemolysis, and feed cells.
A classical one is saline-adenine-glucose-mannitol (SAGM) solution. Also adenosine-citrate -dextrose in Nacl 9°/°° medium (you an find corresponding references on web)
For RBC, you need anticoagulant (citrate) ; adenosine is important , and also dextrose to "feed" cells.
During hematological analysis, total blood is used directly, becase, indeed, centrifugation increases hemolysis rate. So, your speed for centrifugation depends on your RBC isolation procedure, and from which tissue you isolate them. You have to find an equilibrium because if you have some platelets that you want to exlude, if cannot centruge to slowly, (1000 rpm), because you could have some platelet contaminant.
Regards
Didier J
Am very thankful to you all for your valuable replies..
Regards...
Jyothi
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