I applied this method to bird plasma samples.
I loaded the plate image to be more clearer.
In A, B, and C rows there is the same sample in three different amount;
In D and E rows there is another sample in two different amounts. F row is empty because of the small plasma amount available for this sample;
In G and H rows there are the two samples pre-heated before titatrion, in order to inactivate the complement activity.
I can see the difference between wells with agglutination (Ag) (a sort of mat in the well bottom) and not agglutination (a red point).
I do not understand why in column 12 it looks like there is Ag (because it is similar to other columns), even if it is the negative control (just PBS and Rabbit RBCs). The point is, what situation is the right one between:
- there is no Ag even in the previous columns
or
- it happened something that we see Ag in column 12 as well.
Thank you very much in advance to whom will help me.
Being the negative control, I'm sure there sjould not be anything, no hagglutination, even though you use rabbit al the way. H and G are more or less the same even though it is diluted( less each time). I reckon they are serial dilutions. F is empty. Is number 11 marked for some reason.? There is less each time but is similar to previous wells. (maybe you were expecting to be F?) Ask yourself what do the wells contain and if there is a mix of samples.
Hello, Angeles Zorrilla Lopez-Perea,
Thank you very much for your answer.
I made the plate in the pic with my samples and I know what the wells contain.
The problem is that I don't see an evident difference, for example in row D, between well D12 and wells from D5 up to D11.
Therefore, considering that the well D12 is the negative ctrl and the other wells of the same row are similar to it, maybe there is no Ag also in the other wells, but if that is true I should expect a dripping in the wells (rabbit RBCs deposited), especially in the negative ctrl (like this that you can see in row G and H).
Column 11 is marked because the serial dilution ends there.
Thank you again
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