We have encountered an issue with the serum of mice being hemolyzed after the process of centrifugation. Could you please provide guidance to prevent hemolysis ?
Carefully follow the all instructions like cardiac puncture technique, blood collection, post collection handling, centrifugation and all other procedures. is the only way.
Possibly there is some lysis with a fine needle so do not apply too much suction and take the sample slowly. Check that you are not centrifuging at too high a speed and crushing the red cells....possibly lower g forcr and longer spin. If your animal house has trained technicians taking blood then speak to them about how they do it
Vandna Bhardwaj can you please explain more how you handle the blood sample.like needle size..?
Because RBCs got lysis not serum...
so
when you take blood sample out the syringe, remove the needle then take out the blood to tubes.
if you need serum than do not centrifuge, just keep at room temperature or 4C For one day.
if you need plasma then do not use more then 4000rpm.
An afterthought. When you are expressing the blood into the collection tube remove the needle so that the blood flows easily from the syringe and not through the needle again.. Spin the cells down at 1200 rpm or even less to avoid stressing the cells
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