I want to examine neutrophil populations in whole blood. I've read that dot plots of CD45 and SSC can be used to gate for granulocytes, but I need to exclude basophils and eosinophils from my analysis. Some papers say that this approach will yield pure neutrophils but others say that other granulocytes will be present.
How can I be sure that I'm only looking at neutrophils?
Hi, Martin.
I think that you will see and granulocytes.
Briefly on neutrophils:
Dimensions in neutrophil morphology smear about 12 microns at the exit from the circulation of about 20 microns. In the bloodstream are about 8-12 hours, about 8 days tissues.
Kernels 2. The core consists of several (usually 3-5) connected with each other segments.). But there are stab neutrophils, their core is not segmented, and has the shape of a curved stick.
The granules in the cytoplasm - are specific fine granules, which are painted in purple-pink color. a) The neutrophil granules are missing lysosomal enzymes, but contains substances of antibacterial action - other lysozyme c) After phagocytosis of bacteria -vnachale merge with the phagosome secondary granules and kill bacteria; -Then merge primary granules (with lysosomal enzymes) and digest the contents.
Organelles contain few mitochondria, a large amount of glycogen, organelles responsible for the synthesis of substances are practically absent.
Functions a) Neutrophils are phagocytes: migrate from the blood and other tissues are phagocytose bacteria and other particles which can lead to local inflammatory reactions.
Granulocytes:
a) Each type contains a type of granulocyte specific granules. b) The latter, depending on the type of cells that have neutrophilic, basophilic or eosinophilic properties. c) It defines possible functional cells in general.
Maximum net population of neutrophils is, if you can eliminate the hydrolytic enzymes granulocytes, including acid phosphatase.
Otherwise, granulocytes will be present. And if the donor blood with inflammation, granulocyte titration will be high.
(Private opinion, flow cytometry did not check)
Regards
You can get then by gating CD16+ CCR3-. please have a look at "CD48 on blood leukocytes and in serum of asthma patients varies with severity, allergy, 2016. If you do not have access, please let me know, I will send u the full text.
Good Luck.
Dear Hyde Chen.
Perhaps you are right, and that is exactly what is needed for the purity of the experiment. We are talking about the purity of the neutrophil granulocytes and possible presence.
CD66b - may be,because they expressed on granulocytes, to induce activation in neutrophils and to be involved in heterophilic adhesion with CD66c.
But CD15 mediates phagocytosis and chemotaxis of neutrophils. Cause neutrophil. Add your answer chemotaxis protein fragments resulting from the activation of complement, fibrinolytic factors and kinin systems and products leukocyte, and platelet-derived bacterial origin.
Since we are talking about a whole blood as a result of the presence of granulocytes is detected with a high degree of probability
BR
Hi Petrov,
@A.V. Petrov sorry, China have web limited, so it is hard to update the question.
I do not get the point. "Since we are talking about a whole blood as a result of the presence of granulocytes is detected with a high degree of probability" ??what does this mean? Do you mean cd15 and cd66b used for the purity of neutrophil in granulocytes, while your topic is about the purity of neutrophil in whole blood? No marker can identification the neutrophil from whole blood?
Dear Martin,
We published data (paper attached) that could help you to exclude eosinophils as CD157-FLAER+ from CD45+CD33+ peripheral granulocytes. Regarding basophils exclusion I have no direct experience on it, but this other paper attached could be helpfull as well.
Simone
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