Hello Yolanda Zhao

If you may want to use Histopaque for isolating neutrophils, add 3 ml of Histopaque 1119 in a 15-ml conical centrifuge tube. Overlay with 3 ml of Histopaque 1077. Do not disturb the interface between Histopaque 1119 and Histopaque 1077. Overlay the bone marrow cell suspension on top of the Histopaque 1077 layer. Avoid disturbing the interface between the cell suspension and Histopaque 1077. Centrifuge for 30 minutes at 872 × g at room temperature without brake. Collect the neutrophils at the interface of the Histopaque 1119 and Histopaque 1077 layers.

For more information, you may want to refer to the article attached below.

Article Isolation of Mouse Neutrophils

If you may want to use Percoll for isolating neutrophils, the cells may be treated on a three-layer Percoll gradient of 78%, 69%, and 52% Percoll respectively, diluted in HBSS, and centrifuged (1500 g, 30 min, room temperature) without braking. The neutrophils from the 69%/78% interface and the upper part of the 78% layer may be harvested into 1% BSA-coated tubes.

For more information and diagrammatic representation, you may want to refer to the articles attached below.

Article Mouse bone marrow contains large numbers of functionally com...

Chapter Isolation and Characterization of Mouse Neutrophils

Best wishes!

Thank you so much for answering my question. Recently I also tried to isolate neutrophils from bone marrow cells. After centrifugation, I cannot find the correct layer to extract the neutrophils. There are three transparent layers in total, and two interface layer, should I extract the cell layer atop of the 1099 layer?

Thank you so much for your answer! Malcolm Nobre

Hello Professor Zhao,

Perhaps genetically engineered somatic mutations would solve the issue if you seek more recessive traits and or genes: Code If both the traits and genes desired in genetic engineering ...