help me - (Jul/04/2010 )

Dear freinds </span>
I have a problem is not being able to get a clear serum and free of any coulor( red) which may be the result analyzing the blood when it is withdrawn from the eyes of mice infected with toxoplasmosis and taken medicine for two months and the serum not clear cause of error in my results </span> in my performance an</span>d for measuring enzymes and now I am a very big problem .please help me get out as quickly as possible, and that I can</span> </span>

i have problem in getting clear serum from mice blood. i collect the blood from orbital sinus (eye), keep it at 37 C for 1 hr. then keep it in fridge (4 degree C) for 3-4 hrs. but when i centrifuge it, blood gets hemolysed. this prob doesn't occur with rabbit blood. whats the reason for the hemolysis, because i handle the sample very gently? please help..

Hi Ismail,
I was having same problem like you before. Same like you, the problem doesn't happen to rabbit, horse, sheep and human blood when I was working with them. I haven't solve the problem yet (as I had other project in hand now), but I do think maybe is due to the osmotic pressure in our buffer (not isotonic enough to the cells). I was thinking of increase the salt concentration in the washing buffer, but I haven't try it yet as I had lack of time.

Anybody got similar experiences or advices?

Adrian.
P/S: my washing and suspension buffer is PBS.

wrong reading.

My friends
Do you know anyone who can help me solve this problem ?

Hello Muna,

I sometimes have had this problem with bovine and sheep blood. The samples tended to be more hemolysed if the draw was a difficult draw (i.e., the animal was very stressed or the needle had to be moved several times before finding the vein).

Have you tried placing the samples on ice right away? This may also decrease hemolysis.

Regards,

lab rat

Hi lab rat
Thank you very much for assistance, but the problem now that you have to withdraw blood from the eye and not from a vein. and there was degradation in serum drawn, when i began using the serum that not clear (mean red serum) to read my enzymes were my results above, I think there is an error please help my as u could and tell my
How can I get rid of my reading and I hope the mistakes that can provide me with a call to ask which is the dissolved amount of blood mixed with serum to be reading is correct. and Is this the right way can i use it? to make my results correct. please i need the answer as fast as u could because i have no time to repeat my work hhhhhhhhhhhhhhhhhhhhhhhhhhhhhhheeeeeeeeeeeeeeeeeeeeeeeeeeeeellllllllllllllllllll
lllllp me my friend


regards, </span></span>

Hi muna,
Found an article:
"Seroprevalence of Toxoplasma gondii Antibodies in Wild Dolphins From the Spanish Mediterranean Coast" www.seaturtle.org/PDF/Cabezon_2004_JParasitol.pdf

The group after withdraw the blood did not directly centrifuge it. Instead, they tried dilutions (should be using some sort of isotonic solutions) and filtration.

My concern is rather the method when you both withdraw and release blood from the syringe that might contributes to the hemolysis. There is articles about retro-orbital bleeding....not sure whether it helps... http://www.theodora.com/rodent_laboratory/...eline_00_1.html

BTW, Did Toxoplasma gondii causes hemolysis?