Quantification of Brain Immune Cells

Mice were transcardially perfused with PBS and brains were removed and subjected to enzymatic digestion using the Neural Tissue Dissociation Kit (P) (Miltenyi Biotec, Auburn, CA). Myelin debris was removed with myelin removal beads II (Miltenyi) and cells were labeled with fluorescence-labeled CD11b (M1/70) (Biolegend, #101218), CD45 (30-F11) (BD # 553079), and Ly6G (Biolegend, #127618) antibodies. Microglia (CD11b^highCD45^med), macrophages (CD11b^highCD45^highLy6G^low), neutrophils (CD11b^highCD45^highLy6G^high), and lymphocytes (CD11b^lowCD45^high) were quantitated by FACS as previously described [29 (link)].

Free full text: Click here

Wu L., Chung J.Y., Cao T., Jin G., Edmiston WJ I.I.I., Hickman S., Levy E.S., Whalen J.A., Abrams E.S., Degterev A., Lo E.H., Tozzi L., Kaplan D.L., El Khoury J, & Whalen M.J. (2021). Genetic inhibition of RIPK3 ameliorates functional outcome in controlled cortical impact independent of necroptosis. Cell Death & Disease, 12(11), 1064.

Publication 2021

Neural Tissue Dissociation Kit (P) myelin removal beads II CD11b (M1/70) CD45 (30-F11)

Antibodies Brains Cd11b Cells Digestion Enzymatic Fluorescence Lymphocytes Macrophages Mice Microglia Myelin Neural tissue Neutrophils

Corresponding Organization :

Other organizations : Massachusetts General Hospital, Harvard University, Tufts University

Top 5 similar protocols

Variable analysis

independent variables

Enzymatic digestion using the Neural Tissue Dissociation Kit (P) (Miltenyi Biotec, Auburn, CA)

dependent variables

Quantification of microglia (CD11b^high CD45^med), macrophages (CD11b^high CD45^high Ly6G^low), neutrophils (CD11b^high CD45^high Ly6G^high), and lymphocytes (CD11b^low CD45^high) by FACS

control variables

Transcardial perfusion with PBS

controls

No positive or negative controls were explicitly mentioned in the given information.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!